Hardware requirements and performance benchmark

Summary

For the preprocessing step, the rnaends package makes use of fastqStreamer from the ShortRead package. Thus, the yieldSize parameter allows to reduce memory requirement as only a certain number of reads will be processed at a time (default is set to 1 million in rnaends).

For the mapping of RNA-Seq reads to the reference sequences, the rnaends package simply uses on the wrappers Rbowtie and Rsubread, respectively for the bowtie and subread aligners.

For quantification step, the count table (internally stored as a tibble), and the downstream analyses, the memory needed depends mainly on the genome or reference sequences studied (i.e. the number of positions corresponding to 5’ and/or 3’ ends observed in the RNA-Seq reads). Thus, the memory needed scales linearly with the length of the reference sequences and the number of experiments/samples/conditions.

Note

Conclusion: All the notebooks presented in the vignettes, and this benchmark can run on a laptop with 16GB memory.

The running times and memory requirements scale linearly

  • with the number of sequencing reads for the pre-processing steps and their mapping to references sequences,

  • with the number of 5’/3’ end locations and samples in the count table.

Material and methods

Hardware: The following was performed on a Dell Latitude 5280 laptop with dual core CPU Intel i5-7300 with 16GB memory.

The package peakRAM was chosen as it intends to capture real memory requirements within R, whereas others generally only capture memory allocations from R functions and data structures (but not compiled code). From the authors: «So-called “copy-on-modify” behavior, characteristic of R, means that some expressions or functions may require an unexpectedly large amount of RAM overhead. For example, replacing a single value in a matrix (e.g., with ‘[<-’) duplicates that matrix in the backend, making this task require twice as much RAM as that used by the matrix itself.»

library(pryr)
library(peakRAM)
library(tidyverse)
library(ggpubr)
mem_used()
#> 90.9 MB
library(rnaends)
mem_used()
#> 813 MB

Dataset

FASTQ files with variable sequencing depths (1 to 20 millions of reads per experimental condition) were generated based on observed distributions of 5’ ends in two conditions (5’PPP only RNAs vs. background noise as in the TSS-EMOTE protocol) to evaluate the performances of different features of the rnaends package:

  • pre-processing of raw reads

    • only validate then extract the mappable part of reads.

    • checking the presence and validity of some sequences, i.e. a recognition sequence (CGGCACCAACCGAGG) and a control sequence (CGC) at precise locations on the reads (see the read structure below for details).

    • checking the validity of UMIs and extracting them for further use for PCR duplicates removal.

    • using barcodes for demultiplexing experimental conditions.

  • quantification

    • mapping

    • count table

  • downstream

    • TSS

100 nucleotide long reads were generated.

UMIs are 17 nucleotides long and are composed of only A, C and G.

12 barcodes were used corresponding to two experimental conditions with 3 or 6 replicates depending on the fact that the first 2 or 3 nucleotides of the barcodes are used:

  • AAxx, ACxx, ATxx for 3 replicates of the 5’PPP RNAs condition, and AAAx, AACx, ACAx, ACCx, ATAx, ATCx for the 6 replicates

  • CAxx, CCxx, CTxx for the 3 replicates of the background noise condition, and CAAx, CACx, CCAx, CCCx, CTAx, CTCx for the 6 replicates.

As a result, benchmarks for 12 barcodes process 12 times 1-5-10-15-20 million reads per barcode/condition, and benchmarks with 6 barcodes process 2-10-20-30-40 million reads per barcode/condition. Thus, the biggest analysis presented here (6 barcodes with 20 millions reads per condition), consists in 240 million reads.

Dataset availability: FASTQ files used in this benchmark are available on zenodo at https://doi.org/10.5281/zenodo.17592454

Pre-processing of reads

Reads Parsing and Validation: parse_reads

The reads in the raw FASTQ files have the following read structure:

      1       2     5   6            20     21     37     38 40     41                 100
      N     - barcode - CGGCACCAACCGAGG    - VVVVVVV -     CGC     - XXXXXXXXXXXXXXXXXXXXX
1 random nt - barcode - recognition.seq    -   UMI   - control.seq -     RNA 5' end

FASTQ files with 1 to 20 millions reads:

fq_file_list <- c("prepro.UMI-RC-CS.1M.fq.gz",
                  "prepro.UMI-RC-CS.5M.fq.gz",
                  "prepro.UMI-RC-CS.10M.fq.gz",
                  "prepro.UMI-RC-CS.15M.fq.gz",
                  "prepro.UMI-RC-CS.20M.fq.gz"
                  )

Read features only with a mappable part to validate and extract: parse_reads

Read features with only the mappable part to extract

rfwo <- init_read_features(start=41, width=60)
rfwo %>%
  knitr::kable()

name

start

width

pattern_type

pattern

max_mismatch

readid_prepend

readseq

41

60

nucleotides

ACTG

0

FALSE

Measure time and maximum RAM in use for the parse_read function:

prwo <- sapply(fq_file_list, function(i) peakRAM(
  parse_reads(fastq_file  = i,
  features = rfwo,
  force = TRUE)
  )
)
prwo <- prwo %>%
  t %>%
  as_tibble() %>%
  select(time=Elapsed_Time_sec, 
         peak_ram=Peak_RAM_Used_MiB) %>%
  mutate(time=duration(as.numeric(time), units="seconds"), 
         peak_ram = as.numeric(peak_ram),
         nb_reads = c(1, 5, 10, 15, 20),
         features = "none")
prwo %>%
  knitr::kable()

time

peak_ram

nb_reads

features

27.355s

470

1

none

127.94s (~2.13 minutes)

907

5

none

253.563s (~4.23 minutes)

1010

10

none

378.18s (~6.3 minutes)

1086

15

none

505.403s (~8.42 minutes)

1202

20

none

Read features with a recognition sequence and a control sequence: parse_reads

Read features with a recognition sequence and a control sequence:

rfw_rs <- add_read_feature(rfwo,
                          name="recog",
                          start=6,
                          width=15,
                          pattern = "CGGCACCAACCGAGG",
                          pattern_type = "constant")
rfw_rs <- add_read_feature(rfw_rs,
                          name="ctrl",
                          start=38,
                          width=3,
                          pattern = "CGC",
                          pattern_type = "constant")
rfw_rs %>%
  knitr::kable()

name

start

width

pattern_type

pattern

max_mismatch

readid_prepend

readseq

41

60

nucleotides

ACTG

0

FALSE

recog

6

15

constant

CGGCACCA….

0

FALSE

ctrl

38

3

constant

CGC

0

FALSE

Measure time and maximum RAM in use for the parse_read function:

prw_rs <- sapply(fq_file_list, function(i) peakRAM(
  parse_reads(fastq_file  = i,
  features = rfw_rs,
  force = TRUE)
  )
)
prw_rs <- prw_rs %>%
  t %>%
  as_tibble() %>%
  select(time=Elapsed_Time_sec, 
         peak_ram=Peak_RAM_Used_MiB) %>%
  mutate(time=duration(as.numeric(time), units="seconds"), 
         peak_ram = as.numeric(peak_ram),
         nb_reads = c(1, 5, 10, 15, 20),
         features = "recognition and control seq")
prw_rs %>%
  knitr::kable()

time

peak_ram

nb_reads

features

25.4450000000002s

526

1

recognition and control seq

128.704s (~2.15 minutes)

1247

5

recognition and control seq

255.521s (~4.26 minutes)

1086

10

recognition and control seq

384.678s (~6.41 minutes)

1166

15

recognition and control seq

545.71s (~9.1 minutes)

1046

20

recognition and control seq

Read features with UMIs: parse_reads

Read features with UMIs:

rfw_umi <- add_read_feature(rfwo,
                        name="UMI",
                        start=21,
                        width=17,
                        pattern="ACG",
                        pattern_type="nucleotides",
                        readid_prepend = TRUE)
rfw_umi %>%
  knitr::kable()

name

start

width

pattern_type

pattern

max_mismatch

readid_prepend

readseq

41

60

nucleotides

ACTG

0

FALSE

UMI

21

17

nucleotides

ACG

0

TRUE

Measure time and maximum RAM in use for the parse_read function:

prw_umi <- sapply(fq_file_list, function(i) peakRAM(
  parse_reads(fastq_file  = i,
  features = rfw_umi,
  force = TRUE)
  )
)
prw_umi <- prw_umi %>%
  t %>%
  as_tibble() %>%
  select(time=Elapsed_Time_sec, 
         peak_ram=Peak_RAM_Used_MiB) %>%
  mutate(time=duration(as.numeric(time), units="seconds"), 
         peak_ram = as.numeric(peak_ram),
         nb_reads = c(1, 5, 10, 15, 20),
         features = "UMI")

prw_umi %>%
  knitr::kable()  

time

peak_ram

nb_reads

features

33.8269999999998s

636

1

UMI

167.811s (~2.8 minutes)

1030

5

UMI

343.696s (~5.73 minutes)

1110

10

UMI

514.835s (~8.58 minutes)

1192

15

UMI

669.22s (~11.15 minutes)

1280

20

UMI

Summary

pr <- bind_rows(prwo, prw_rs, prw_umi)

Running time and memory footprint depending on the number of reads:

p1 <- pr %>%
  ggplot() +
  geom_line(aes(x=nb_reads, y=time, color=features),
            show.legend = FALSE) +
  geom_point(aes(x=nb_reads, y=time, shape=features, color=features),
             show.legend = FALSE) +
  theme_light() +
  xlab("Number of reads (in millions)") +
  ylab("Duration (in seconds)") +
  ggtitle("Running times for pre-processing functions") 
p2 <- pr %>%
  ggplot() +
  geom_line(aes(x=nb_reads, y=peak_ram, color=features)) +
  geom_point(aes(x=nb_reads, y=peak_ram, shape=features, color=features)) +
  theme_light() +
  xlab("Number of reads (in millions)") +
  ylab("Memory used (in MB)") +
  ggtitle("Memory used during pre-processing functions") +
  theme(legend.position="inside", legend.position.inside = c(0.6, 0.15))
ggarrange(p1, p2) 

Note

In the above graphs, we can see that the running time scales linearly with the number of sequencing reads to analyse. The overhead for the validation of constant strings at certain locations in reads (i.e. recognition and control sequences) is negligible. The UMI validation and extraction for further analyses scale also linearly with a substantial overhead.

Regarding memory footprint, we can observe that, except for small amounts of reads (below 5 millions), the memory in use is aroung 1-1.5GB. This is to be correlated with the yieldSize parameter (default: 1 million) used by the fastqStreamer from the ShortRead package for processing reads, i.e. reads are loaded and processed by groups of 1 million reads from the FASTQ file, thus only a certain amount of memory is necessary.

Reads Parsing and demultiplexing: demultiplex

Read features with barcodes for demultiplexing: demultiplex

FASTQ files composed of multiplexed FASTQ files with various sequencing depths. 1M means 1 million reads per condition, thus 12 millions reads in total:

fq_files_bc <- c("prepro.PPP-BG.1M.fq.gz", 
                 "prepro.PPP-BG.5M.fq.gz",
                 "prepro.PPP-BG.10M.fq.gz",
                 "prepro.PPP-BG.15M.fq.gz",
                 "prepro.PPP-BG.20M.fq.gz"
                 )

Read features with barcodes of length 2 and 6 experiments: 3 replicates for control samples (background noise), and 3 replicates for test samples (5’PPP samples)

rfw_bc2 <- add_read_feature(rfwo,
                          name="barcode",
                          start=2,
                          width=2,
                          pattern=c("AA","AC","AT","CA","CC","CT"),
                          pattern_type = "constant",
                          readid_prepend = FALSE
)
rfw_bc2 %>%
  knitr::kable()

name

start

width

pattern_type

pattern

max_mismatch

readid_prepend

readseq

41

60

nucleotides

ACTG

0

FALSE

barcode

2

2

constant

AA, AC, ….

0

FALSE

Measure time and maximum RAM in use for the demultiplex function for 6 barcodes:

prw_bc2 <- sapply(fq_files_bc, function(i) peakRAM(
  demultiplex(fastq_file  = i,
              features = rfw_bc2,
              force = TRUE)
  )
)
prw_bc2 <- prw_bc2 %>%
  t %>%
  as_tibble() %>%
  select(time=Elapsed_Time_sec, 
         peak_ram=Peak_RAM_Used_MiB) %>%
  mutate(time=duration(as.numeric(time), units="seconds"), 
         peak_ram = as.numeric(peak_ram),
         nb_reads = c(12, 60, 120, 180, 240),
         features = "6 barcodes")
prw_bc2 %>%
  knitr::kable()

time

peak_ram

nb_reads

features

321.641000000001s (~5.36 minutes)

1098

12

6 barcodes

1602.599s (~26.71 minutes)

1182

60

6 barcodes

4291.41s (~1.19 hours)

1035

120

6 barcodes

4720.811s (~1.31 hours)

1004

180

6 barcodes

6297.646s (~1.75 hours)

1004

240

6 barcodes

Read features with barcodes of length 3 and 12 experiments: 6 replicates for control samples (background noise), and 6 replicates for test samples (5’PPP samples)

rfw_bc3 <- add_read_feature(rfwo,
                            name="barcode",
                            start=2,
                            width=3,
                            pattern=c("AAA","AAC","ACA","ACC","ATA","ATC",
                                      "CAA","CAC","CCA","CCC","CTA","CTC"),
                            pattern_type = "constant",
                            readid_prepend = FALSE
)
rfw_bc3 %>%
  knitr::kable()

name

start

width

pattern_type

pattern

max_mismatch

readid_prepend

readseq

41

60

nucleotides

ACTG

0

FALSE

barcode

2

3

constant

AAA, AAC….

0

FALSE

Measure time and maximum RAM in use for the demultiplex function for 12 barcodes:

prw_bc3 <- sapply(fq_files_bc, function(i) peakRAM(
  demultiplex(fastq_file  = i,
              features = rfw_bc3,
              force = TRUE)
  )
)
prw_bc3 <- prw_bc3 %>%
  t %>%
  as_tibble() %>%
  select(time=Elapsed_Time_sec, 
         peak_ram=Peak_RAM_Used_MiB) %>%
  mutate(time=duration(as.numeric(time), units="seconds"), 
         peak_ram = as.numeric(peak_ram),
         nb_reads = c(12, 60, 120, 180, 240),
         features = "12 barcodes")
prw_bc3 %>%
  knitr::kable()

time

peak_ram

nb_reads

features

316.102999999999s (~5.27 minutes)

1005

12

12 barcodes

1589.093s (~26.48 minutes)

1005

60

12 barcodes

3158.782s (~52.65 minutes)

1247

120

12 barcodes

4731.018s (~1.31 hours)

1084

180

12 barcodes

6256.014s (~1.74 hours)

1165

240

12 barcodes

Read features combined: demultiplex

Read features with recognition and control sequences, UMIs, and barcodes:

rfw_a <- add_read_feature(rfwo,
                          name="recog",
                          start=6,
                          width=15,
                          pattern = "CGGCACCAACCGAGG",
                          pattern_type = "constant")
rfw_a <- add_read_feature(rfw_a,
                          name="ctrl",
                          start=38,
                          width=3,
                          pattern = "CGC",
                          pattern_type = "constant")
rfw_a <- add_read_feature(rfw_a,
                         name="UMI",
                         start=21,
                         width=17,
                         pattern="ACG",
                         pattern_type="nucleotides",
                         readid_prepend = TRUE)
rfw_a <- add_read_feature(rfw_a,
                          name="barcode",
                          start=2,
                          width=2,
                          pattern=c("AA","AC","AT","CA","CC","CT"),
                          pattern_type = "constant",
                          readid_prepend = FALSE)
rfw_a %>%
  knitr::kable()

name

start

width

pattern_type

pattern

max_mismatch

readid_prepend

readseq

41

60

nucleotides

ACTG

0

FALSE

recog

6

15

constant

CGGCACCA….

0

FALSE

ctrl

38

3

constant

CGC

0

FALSE

UMI

21

17

nucleotides

ACG

0

TRUE

barcode

2

2

constant

AA, AC, ….

0

FALSE

Measure time and maximum RAM in use for the demultiplex function for combined read features:

prw_a <- sapply(fq_files_bc, function(i) peakRAM(
  demultiplex(fastq_file  = i,
              features = rfw_a,
              force = TRUE)
  )
)
prw_a <- prw_a %>%
  t %>%
  as_tibble() %>%
  select(time=Elapsed_Time_sec, 
         peak_ram=Peak_RAM_Used_MiB) %>%
  mutate(time=duration(as.numeric(time), units="seconds"), 
         peak_ram = as.numeric(peak_ram),
        nb_reads = c(12, 60, 120, 180, 240),
         features = "RS+CS, UMI, 6 barcodes")
prw_a %>%
  knitr::kable()

time

peak_ram

nb_reads

features

401.449999999997s (~6.69 minutes)

1252

12

RS+CS, UMI, 6 barcodes

1992.30899999999s (~33.21 minutes)

1344

60

RS+CS, UMI, 6 barcodes

3932.47199999999s (~1.09 hours)

1448

120

RS+CS, UMI, 6 barcodes

5881.689s (~1.63 hours)

1238

180

RS+CS, UMI, 6 barcodes

7933s (~2.2 hours)

1355

240

RS+CS, UMI, 6 barcodes

Summary

pr2 <- bind_rows(prw_bc2, prw_bc3, prw_a)

Running time and memory footprint depending on the number of reads:

p1 <- pr2 %>%
  ggplot() +
  geom_line(aes(x=nb_reads, y=time, color=features)) +
  geom_point(aes(x=nb_reads, y=time, shape=features, color=features)) +
  theme_light() +
  xlab("Number of reads (in millions)") +
  ylab("Duration (in seconds)") +
  ggtitle("Running times for demultipexing")  +
  theme(legend.position="inside", legend.position.inside = c(0.6, 0.15))

p2 <- pr2 %>%
  ggplot() +
  geom_line(aes(x=nb_reads, y=peak_ram, color=features),
            show.legend = FALSE) +
  geom_point(aes(x=nb_reads, y=peak_ram, shape=features, color=features),
            show.legend = FALSE) +
  theme_light() +
  xlab("Number of reads (in millions)") +
  ylab("Memory used (in MB)") +
  ggtitle("Memory used during demultipexing")
ggarrange(p1, p2) 

Note

In the above graphs, we can see that the running times for demultiplexing scale linearly with the number of reads to analyse, regardless of the number of analyses to demultiplex. As in the previous section, the overhead of additional features to parse, validate and extract (recogntion sequence denoted RS, control sequence denoted CS, and UMIs) scale also linaerly with the size of the input.

Regarding the memory footprint, we observe the same behavior as in the previous section, i.e. it mainly depends on the yieldSize parameter used for processing reads per chunk by the fastqStreamer from the ShortRead package.

Quantification

Data preparation: use previous read features with combined features.

dirout <- c("quantif.1M", "quantif.5M", "quantif.10M", "quantif.15M", "quantif.20M")
names(dirout) <- fq_files_bc
sapply(fq_files_bc, function(i)
  demultiplex(fastq_file  = i,
              features = rfw_a,
              out_dir = dirout[i],
              force = FALSE)
)
#>                  prepro.PPP-BG.1M.fq.gz prepro.PPP-BG.5M.fq.gz
#> barcode_group    character,6            character,6           
#> is_valid_readseq logical,6              logical,6             
#> is_valid_recog   logical,6              logical,6             
#> is_valid_ctrl    logical,6              logical,6             
#> is_valid_UMI     logical,6              logical,6             
#> is_valid_barcode logical,6              logical,6             
#> count            integer,6              integer,6             
#> demux_filename   character,6            character,6           
#>                  prepro.PPP-BG.10M.fq.gz prepro.PPP-BG.15M.fq.gz
#> barcode_group    character,6             character,6            
#> is_valid_readseq logical,6               logical,6              
#> is_valid_recog   logical,6               logical,6              
#> is_valid_ctrl    logical,6               logical,6              
#> is_valid_UMI     logical,6               logical,6              
#> is_valid_barcode logical,6               logical,6              
#> count            integer,6               integer,6              
#> demux_filename   character,6             character,6            
#>                  prepro.PPP-BG.20M.fq.gz
#> barcode_group    character,6            
#> is_valid_readseq logical,6              
#> is_valid_recog   logical,6              
#> is_valid_ctrl    logical,6              
#> is_valid_UMI     logical,6              
#> is_valid_barcode logical,6              
#> count            integer,6              
#> demux_filename   character,6

Mapping: map_fastq_files

Measure time and maximum RAM in use for the map_fastq_files function with bowtie:

prw_bt <- sapply(dirout, function(i) peakRAM(
  map_fastq_files(
    list_fastq_files = list.files(i,
                                  pattern = "_valid.fastq.gz$",
                                  full.names = TRUE),
    index_name = "MW2.bt",
    fasta_name = "MW2.fasta",
    max_map_mismatch = 1,
    aligner = "bowtie",
    map_unique = TRUE,
    force = TRUE
  ))
)
#> Warning in FUN(X[[i]], ...): Index directory already exists remove it if you
#> want to recreate it
#> Warning in FUN(X[[i]], ...): Index directory already exists remove it if you
#> want to recreate it
#> Warning in FUN(X[[i]], ...): Index directory already exists remove it if you
#> want to recreate it
#> Warning in FUN(X[[i]], ...): Index directory already exists remove it if you
#> want to recreate it
#> Warning in FUN(X[[i]], ...): Index directory already exists remove it if you
#> want to recreate it
#> Warning in FUN(X[[i]], ...): Index directory already exists remove it if you
#> want to recreate it
#> Warning in FUN(X[[i]], ...): Index directory already exists remove it if you
#> want to recreate it
#> Warning in FUN(X[[i]], ...): Index directory already exists remove it if you
#> want to recreate it
#> Warning in FUN(X[[i]], ...): Index directory already exists remove it if you
#> want to recreate it
#> Warning in FUN(X[[i]], ...): Index directory already exists remove it if you
#> want to recreate it
#> Warning in FUN(X[[i]], ...): Index directory already exists remove it if you
#> want to recreate it
#> Warning in FUN(X[[i]], ...): Index directory already exists remove it if you
#> want to recreate it
#> Warning in FUN(X[[i]], ...): Index directory already exists remove it if you
#> want to recreate it
#> Warning in FUN(X[[i]], ...): Index directory already exists remove it if you
#> want to recreate it
#> Warning in FUN(X[[i]], ...): Index directory already exists remove it if you
#> want to recreate it
#> Warning in FUN(X[[i]], ...): Index directory already exists remove it if you
#> want to recreate it
#> Warning in FUN(X[[i]], ...): Index directory already exists remove it if you
#> want to recreate it
#> Warning in FUN(X[[i]], ...): Index directory already exists remove it if you
#> want to recreate it
#> Warning in FUN(X[[i]], ...): Index directory already exists remove it if you
#> want to recreate it
#> Warning in FUN(X[[i]], ...): Index directory already exists remove it if you
#> want to recreate it
#> Warning in FUN(X[[i]], ...): Index directory already exists remove it if you
#> want to recreate it
#> Warning in FUN(X[[i]], ...): Index directory already exists remove it if you
#> want to recreate it
#> Warning in FUN(X[[i]], ...): Index directory already exists remove it if you
#> want to recreate it
#> Warning in FUN(X[[i]], ...): Index directory already exists remove it if you
#> want to recreate it
#> Warning in FUN(X[[i]], ...): Index directory already exists remove it if you
#> want to recreate it
#> Warning in FUN(X[[i]], ...): Index directory already exists remove it if you
#> want to recreate it
#> Warning in FUN(X[[i]], ...): Index directory already exists remove it if you
#> want to recreate it
#> Warning in FUN(X[[i]], ...): Index directory already exists remove it if you
#> want to recreate it
#> Warning in FUN(X[[i]], ...): Index directory already exists remove it if you
#> want to recreate it
#> Warning in FUN(X[[i]], ...): Index directory already exists remove it if you
#> want to recreate it
prw_bt <- prw_bt %>%
  t %>%
  as_tibble() %>%
  select(time=Elapsed_Time_sec, 
         peak_ram=Peak_RAM_Used_MiB) %>%
  mutate(time=duration(as.numeric(time), units="seconds"), 
         peak_ram = as.numeric(peak_ram),
         nb_reads = c(12, 60, 120, 180, 240),
         aligner = "bowtie")
prw_bt %>%
  knitr::kable()

time

peak_ram

nb_reads

features

229.714000000007s (~3.83 minutes)

1.2

12

bowtie

1192.98299999999s (~19.88 minutes)

0.5

60

bowtie

2357.016s (~39.28 minutes)

0.5

120

bowtie

3545.39999999999s (~59.09 minutes)

0.5

180

bowtie

4709.34299999999s (~1.31 hours)

0.5

240

bowtie

Measure time and maximum RAM in use for the map_fastq_files function with subread:

prw_sr <- sapply(dirout, function(i) peakRAM(
  map_fastq_files(
    list_fastq_files = list.files(i,
                                  pattern = "_valid.fastq.gz$",
                                  full.names = TRUE),
    index_name = "MW2.sr",
    fasta_name = "MW2.fasta",
    max_map_mismatch = 1,
    aligner = "subread",
    map_unique = TRUE,
    force = TRUE
  ))
)
subread output
#> [1] "Index directory already exists remove it if you want to recreate it"
#> 
#>         ==========     _____ _    _ ____  _____  ______          _____  
#>         =====         / ____| |  | |  _ \|  __ \|  ____|   /\   |  __ \ 
#>           =====      | (___ | |  | | |_) | |__) | |__     /  \  | |  | |
#>             ====      \___ \| |  | |  _ <|  _  /|  __|   / /\ \ | |  | |
#>               ====    ____) | |__| | |_) | | \ \| |____ / ____ \| |__| |
#>         ==========   |_____/ \____/|____/|_|  \_\______/_/    \_\_____/
#>        Rsubread 2.20.0
#> 
#> //================================= setting ==================================\\
#> ||                                                                            ||
#> || Function      : Read alignment (DNA-Seq)                                   ||
#> || Input file    : prepro.PPP-BG.1M_AA_valid.fastq.gz                         ||
#> || Output file   : prepro.PPP-BG.1M_AA_valid.fastq.gz.bam (BAM), Sorted       ||
#> || Index name    : index                                                      ||
#> ||                                                                            ||
#> ||                    ------------------------------------                    ||
#> ||                                                                            ||
#> ||                               Threads : 1                                  ||
#> ||                          Phred offset : 33                                 ||
#> ||                             Min votes : 3 / 10                             ||
#> ||                        Max mismatches : 1                                  ||
#> ||                      Max indel length : 0                                  ||
#> ||            Report multi-mapping reads : no                                 ||
#> || Max alignments per multi-mapping read : 1                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> //================ Running (01-Nov-2025 17:32:38, pid=392792) ================\\
#> ||                                                                            ||
#> || Check the input reads.                                                     ||
#> || The input file contains base space reads.                                  ||
#> || Initialise the memory objects.                                             ||
#> || Estimate the mean read length.                                             ||
#> || WARNING  - The specified Phred score offset (33) seems incorrect.          ||
#> ||            ASCII values of the quality scores of read bases included in    ||
#> ||            the first 3000 reads were found to be within the range of       ||
#> ||            [63,63].                                                        ||
#> ||                                                                            ||
#> || Create the output BAM file.                                                ||
#> || Check the index.                                                           ||
#> || Init the voting space.                                                     ||
#> || Global environment is initialised.                                         ||
#> || Load the 1-th index block...                                               ||
#> || The index block has been loaded.                                           ||
#> || Start read mapping in chunk.                                               ||
#> ||    0% completed, 1.1 mins elapsed, rate=73.5k reads per second             ||
#> ||    6% completed, 1.2 mins elapsed, rate=102.7k reads per second            ||
#> ||   13% completed, 1.2 mins elapsed, rate=104.6k reads per second            ||
#> ||   19% completed, 1.2 mins elapsed, rate=105.4k reads per second            ||
#> ||   26% completed, 1.2 mins elapsed, rate=105.0k reads per second            ||
#> ||   33% completed, 1.3 mins elapsed, rate=103.9k reads per second            ||
#> ||   39% completed, 1.3 mins elapsed, rate=104.4k reads per second            ||
#> ||   46% completed, 1.3 mins elapsed, rate=104.5k reads per second            ||
#> ||   53% completed, 1.3 mins elapsed, rate=103.4k reads per second            ||
#> ||   59% completed, 1.3 mins elapsed, rate=103.2k reads per second            ||
#> ||   66% completed, 1.4 mins elapsed, rate=103.3k reads per second            ||
#> ||   70% completed, 1.4 mins elapsed, rate=16.9k reads per second             ||
#> ||   73% completed, 1.4 mins elapsed, rate=17.5k reads per second             ||
#> ||   76% completed, 1.4 mins elapsed, rate=18.1k reads per second             ||
#> ||   79% completed, 1.4 mins elapsed, rate=18.7k reads per second             ||
#> ||   83% completed, 1.4 mins elapsed, rate=19.2k reads per second             ||
#> ||   86% completed, 1.5 mins elapsed, rate=19.8k reads per second             ||
#> ||   90% completed, 1.5 mins elapsed, rate=20.4k reads per second             ||
#> ||   93% completed, 1.5 mins elapsed, rate=20.9k reads per second             ||
#> ||   96% completed, 1.5 mins elapsed, rate=21.4k reads per second             ||
#> ||                                                                            ||
#> ||                           Completed successfully.                          ||
#> ||                                                                            ||
#> \\====================================    ====================================//
#> 
#> //================================   Summary =================================\\
#> ||                                                                            ||
#> ||                 Total reads : 2,000,000                                    ||
#> ||                      Mapped : 1,422,829 (71.1%)                            ||
#> ||             Uniquely mapped : 1,422,829                                    ||
#> ||               Multi-mapping : 0                                            ||
#> ||                                                                            ||
#> ||                    Unmapped : 577,171                                      ||
#> ||                                                                            ||
#> ||                      Indels : 0                                            ||
#> ||                                                                            ||
#> ||                     WARNING : Phred offset (33) incorrect?                 ||
#> ||                                                                            ||
#> ||                Running time : 1.6 minutes                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> [1] "Index directory already exists remove it if you want to recreate it"
#> 
#>         ==========     _____ _    _ ____  _____  ______          _____  
#>         =====         / ____| |  | |  _ \|  __ \|  ____|   /\   |  __ \ 
#>           =====      | (___ | |  | | |_) | |__) | |__     /  \  | |  | |
#>             ====      \___ \| |  | |  _ <|  _  /|  __|   / /\ \ | |  | |
#>               ====    ____) | |__| | |_) | | \ \| |____ / ____ \| |__| |
#>         ==========   |_____/ \____/|____/|_|  \_\______/_/    \_\_____/
#>        Rsubread 2.20.0
#> 
#> //================================= setting ==================================\\
#> ||                                                                            ||
#> || Function      : Read alignment (DNA-Seq)                                   ||
#> || Input file    : prepro.PPP-BG.1M_AC_valid.fastq.gz                         ||
#> || Output file   : prepro.PPP-BG.1M_AC_valid.fastq.gz.bam (BAM), Sorted       ||
#> || Index name    : index                                                      ||
#> ||                                                                            ||
#> ||                    ------------------------------------                    ||
#> ||                                                                            ||
#> ||                               Threads : 1                                  ||
#> ||                          Phred offset : 33                                 ||
#> ||                             Min votes : 3 / 10                             ||
#> ||                        Max mismatches : 1                                  ||
#> ||                      Max indel length : 0                                  ||
#> ||            Report multi-mapping reads : no                                 ||
#> || Max alignments per multi-mapping read : 1                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> //================ Running (01-Nov-2025 17:34:13, pid=392792) ================\\
#> ||                                                                            ||
#> || Check the input reads.                                                     ||
#> || The input file contains base space reads.                                  ||
#> || Initialise the memory objects.                                             ||
#> || Estimate the mean read length.                                             ||
#> || WARNING  - The specified Phred score offset (33) seems incorrect.          ||
#> ||            ASCII values of the quality scores of read bases included in    ||
#> ||            the first 3000 reads were found to be within the range of       ||
#> ||            [63,63].                                                        ||
#> ||                                                                            ||
#> || Create the output BAM file.                                                ||
#> || Check the index.                                                           ||
#> || Init the voting space.                                                     ||
#> || Global environment is initialised.                                         ||
#> || Load the 1-th index block...                                               ||
#> || The index block has been loaded.                                           ||
#> || Start read mapping in chunk.                                               ||
#> ||    0% completed, 1.1 mins elapsed, rate=102.4k reads per second            ||
#> ||    6% completed, 1.2 mins elapsed, rate=109.0k reads per second            ||
#> ||   13% completed, 1.2 mins elapsed, rate=109.0k reads per second            ||
#> ||   19% completed, 1.2 mins elapsed, rate=108.8k reads per second            ||
#> ||   26% completed, 1.2 mins elapsed, rate=108.9k reads per second            ||
#> ||   33% completed, 1.2 mins elapsed, rate=108.9k reads per second            ||
#> ||   39% completed, 1.3 mins elapsed, rate=109.1k reads per second            ||
#> ||   46% completed, 1.3 mins elapsed, rate=109.0k reads per second            ||
#> ||   53% completed, 1.3 mins elapsed, rate=108.8k reads per second            ||
#> ||   59% completed, 1.3 mins elapsed, rate=108.7k reads per second            ||
#> ||   66% completed, 1.3 mins elapsed, rate=108.5k reads per second            ||
#> ||   69% completed, 1.4 mins elapsed, rate=17.1k reads per second             ||
#> ||   73% completed, 1.4 mins elapsed, rate=17.7k reads per second             ||
#> ||   76% completed, 1.4 mins elapsed, rate=18.3k reads per second             ||
#> ||   79% completed, 1.4 mins elapsed, rate=18.9k reads per second             ||
#> ||   83% completed, 1.4 mins elapsed, rate=19.4k reads per second             ||
#> ||   86% completed, 1.4 mins elapsed, rate=20.0k reads per second             ||
#> ||   89% completed, 1.5 mins elapsed, rate=20.6k reads per second             ||
#> ||   93% completed, 1.5 mins elapsed, rate=21.1k reads per second             ||
#> ||   96% completed, 1.5 mins elapsed, rate=21.7k reads per second             ||
#> ||                                                                            ||
#> ||                           Completed successfully.                          ||
#> ||                                                                            ||
#> \\====================================    ====================================//
#> 
#> //================================   Summary =================================\\
#> ||                                                                            ||
#> ||                 Total reads : 2,000,000                                    ||
#> ||                      Mapped : 1,423,160 (71.2%)                            ||
#> ||             Uniquely mapped : 1,423,160                                    ||
#> ||               Multi-mapping : 0                                            ||
#> ||                                                                            ||
#> ||                    Unmapped : 576,840                                      ||
#> ||                                                                            ||
#> ||                      Indels : 0                                            ||
#> ||                                                                            ||
#> ||                     WARNING : Phred offset (33) incorrect?                 ||
#> ||                                                                            ||
#> ||                Running time : 1.6 minutes                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> [1] "Index directory already exists remove it if you want to recreate it"
#> 
#>         ==========     _____ _    _ ____  _____  ______          _____  
#>         =====         / ____| |  | |  _ \|  __ \|  ____|   /\   |  __ \ 
#>           =====      | (___ | |  | | |_) | |__) | |__     /  \  | |  | |
#>             ====      \___ \| |  | |  _ <|  _  /|  __|   / /\ \ | |  | |
#>               ====    ____) | |__| | |_) | | \ \| |____ / ____ \| |__| |
#>         ==========   |_____/ \____/|____/|_|  \_\______/_/    \_\_____/
#>        Rsubread 2.20.0
#> 
#> //================================= setting ==================================\\
#> ||                                                                            ||
#> || Function      : Read alignment (DNA-Seq)                                   ||
#> || Input file    : prepro.PPP-BG.1M_AT_valid.fastq.gz                         ||
#> || Output file   : prepro.PPP-BG.1M_AT_valid.fastq.gz.bam (BAM), Sorted       ||
#> || Index name    : index                                                      ||
#> ||                                                                            ||
#> ||                    ------------------------------------                    ||
#> ||                                                                            ||
#> ||                               Threads : 1                                  ||
#> ||                          Phred offset : 33                                 ||
#> ||                             Min votes : 3 / 10                             ||
#> ||                        Max mismatches : 1                                  ||
#> ||                      Max indel length : 0                                  ||
#> ||            Report multi-mapping reads : no                                 ||
#> || Max alignments per multi-mapping read : 1                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> //================ Running (01-Nov-2025 17:35:48, pid=392792) ================\\
#> ||                                                                            ||
#> || Check the input reads.                                                     ||
#> || The input file contains base space reads.                                  ||
#> || Initialise the memory objects.                                             ||
#> || Estimate the mean read length.                                             ||
#> || WARNING  - The specified Phred score offset (33) seems incorrect.          ||
#> ||            ASCII values of the quality scores of read bases included in    ||
#> ||            the first 3000 reads were found to be within the range of       ||
#> ||            [63,63].                                                        ||
#> ||                                                                            ||
#> || Create the output BAM file.                                                ||
#> || Check the index.                                                           ||
#> || Init the voting space.                                                     ||
#> || Global environment is initialised.                                         ||
#> || Load the 1-th index block...                                               ||
#> || The index block has been loaded.                                           ||
#> || Start read mapping in chunk.                                               ||
#> ||    0% completed, 1.2 mins elapsed, rate=99.3k reads per second             ||
#> ||    6% completed, 1.2 mins elapsed, rate=109.3k reads per second            ||
#> ||   13% completed, 1.2 mins elapsed, rate=107.0k reads per second            ||
#> ||   19% completed, 1.2 mins elapsed, rate=107.9k reads per second            ||
#> ||   26% completed, 1.2 mins elapsed, rate=108.6k reads per second            ||
#> ||   32% completed, 1.3 mins elapsed, rate=108.8k reads per second            ||
#> ||   39% completed, 1.3 mins elapsed, rate=108.9k reads per second            ||
#> ||   46% completed, 1.3 mins elapsed, rate=109.0k reads per second            ||
#> ||   52% completed, 1.3 mins elapsed, rate=109.0k reads per second            ||
#> ||   59% completed, 1.3 mins elapsed, rate=109.1k reads per second            ||
#> ||   66% completed, 1.4 mins elapsed, rate=108.6k reads per second            ||
#> ||   70% completed, 1.4 mins elapsed, rate=16.9k reads per second             ||
#> ||   73% completed, 1.4 mins elapsed, rate=17.5k reads per second             ||
#> ||   76% completed, 1.4 mins elapsed, rate=18.2k reads per second             ||
#> ||   79% completed, 1.4 mins elapsed, rate=18.7k reads per second             ||
#> ||   83% completed, 1.4 mins elapsed, rate=19.3k reads per second             ||
#> ||   86% completed, 1.5 mins elapsed, rate=19.9k reads per second             ||
#> ||   89% completed, 1.5 mins elapsed, rate=20.4k reads per second             ||
#> ||   93% completed, 1.5 mins elapsed, rate=21.0k reads per second             ||
#> ||   96% completed, 1.5 mins elapsed, rate=21.5k reads per second             ||
#> ||                                                                            ||
#> ||                           Completed successfully.                          ||
#> ||                                                                            ||
#> \\====================================    ====================================//
#> 
#> //================================   Summary =================================\\
#> ||                                                                            ||
#> ||                 Total reads : 2,000,000                                    ||
#> ||                      Mapped : 1,424,301 (71.2%)                            ||
#> ||             Uniquely mapped : 1,424,301                                    ||
#> ||               Multi-mapping : 0                                            ||
#> ||                                                                            ||
#> ||                    Unmapped : 575,699                                      ||
#> ||                                                                            ||
#> ||                      Indels : 0                                            ||
#> ||                                                                            ||
#> ||                     WARNING : Phred offset (33) incorrect?                 ||
#> ||                                                                            ||
#> ||                Running time : 1.6 minutes                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> [1] "Index directory already exists remove it if you want to recreate it"
#> 
#>         ==========     _____ _    _ ____  _____  ______          _____  
#>         =====         / ____| |  | |  _ \|  __ \|  ____|   /\   |  __ \ 
#>           =====      | (___ | |  | | |_) | |__) | |__     /  \  | |  | |
#>             ====      \___ \| |  | |  _ <|  _  /|  __|   / /\ \ | |  | |
#>               ====    ____) | |__| | |_) | | \ \| |____ / ____ \| |__| |
#>         ==========   |_____/ \____/|____/|_|  \_\______/_/    \_\_____/
#>        Rsubread 2.20.0
#> 
#> //================================= setting ==================================\\
#> ||                                                                            ||
#> || Function      : Read alignment (DNA-Seq)                                   ||
#> || Input file    : prepro.PPP-BG.1M_CA_valid.fastq.gz                         ||
#> || Output file   : prepro.PPP-BG.1M_CA_valid.fastq.gz.bam (BAM), Sorted       ||
#> || Index name    : index                                                      ||
#> ||                                                                            ||
#> ||                    ------------------------------------                    ||
#> ||                                                                            ||
#> ||                               Threads : 1                                  ||
#> ||                          Phred offset : 33                                 ||
#> ||                             Min votes : 3 / 10                             ||
#> ||                        Max mismatches : 1                                  ||
#> ||                      Max indel length : 0                                  ||
#> ||            Report multi-mapping reads : no                                 ||
#> || Max alignments per multi-mapping read : 1                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> //================ Running (01-Nov-2025 17:37:23, pid=392792) ================\\
#> ||                                                                            ||
#> || Check the input reads.                                                     ||
#> || The input file contains base space reads.                                  ||
#> || Initialise the memory objects.                                             ||
#> || Estimate the mean read length.                                             ||
#> || WARNING  - The specified Phred score offset (33) seems incorrect.          ||
#> ||            ASCII values of the quality scores of read bases included in    ||
#> ||            the first 3000 reads were found to be within the range of       ||
#> ||            [63,63].                                                        ||
#> ||                                                                            ||
#> || Create the output BAM file.                                                ||
#> || Check the index.                                                           ||
#> || Init the voting space.                                                     ||
#> || Global environment is initialised.                                         ||
#> || Load the 1-th index block...                                               ||
#> || The index block has been loaded.                                           ||
#> || Start read mapping in chunk.                                               ||
#> ||    0% completed, 1.1 mins elapsed, rate=105.9k reads per second            ||
#> ||    6% completed, 1.2 mins elapsed, rate=114.8k reads per second            ||
#> ||   13% completed, 1.2 mins elapsed, rate=115.5k reads per second            ||
#> ||   20% completed, 1.2 mins elapsed, rate=115.4k reads per second            ||
#> ||   26% completed, 1.2 mins elapsed, rate=115.2k reads per second            ||
#> ||   33% completed, 1.2 mins elapsed, rate=115.0k reads per second            ||
#> ||   40% completed, 1.3 mins elapsed, rate=115.0k reads per second            ||
#> ||   46% completed, 1.3 mins elapsed, rate=115.1k reads per second            ||
#> ||   53% completed, 1.3 mins elapsed, rate=115.1k reads per second            ||
#> ||   60% completed, 1.3 mins elapsed, rate=114.8k reads per second            ||
#> ||   70% completed, 1.4 mins elapsed, rate=17.1k reads per second             ||
#> ||   73% completed, 1.4 mins elapsed, rate=17.7k reads per second             ||
#> ||   76% completed, 1.4 mins elapsed, rate=18.3k reads per second             ||
#> ||   80% completed, 1.4 mins elapsed, rate=18.8k reads per second             ||
#> ||   83% completed, 1.4 mins elapsed, rate=19.4k reads per second             ||
#> ||   86% completed, 1.5 mins elapsed, rate=19.9k reads per second             ||
#> ||   89% completed, 1.5 mins elapsed, rate=20.4k reads per second             ||
#> ||   93% completed, 1.5 mins elapsed, rate=20.9k reads per second             ||
#> ||   96% completed, 1.5 mins elapsed, rate=21.4k reads per second             ||
#> ||                                                                            ||
#> ||                           Completed successfully.                          ||
#> ||                                                                            ||
#> \\====================================    ====================================//
#> 
#> //================================   Summary =================================\\
#> ||                                                                            ||
#> ||                 Total reads : 2,000,000                                    ||
#> ||                      Mapped : 374,571 (18.7%)                              ||
#> ||             Uniquely mapped : 374,571                                      ||
#> ||               Multi-mapping : 0                                            ||
#> ||                                                                            ||
#> ||                    Unmapped : 1,625,429                                    ||
#> ||                                                                            ||
#> ||                      Indels : 0                                            ||
#> ||                                                                            ||
#> ||                     WARNING : Phred offset (33) incorrect?                 ||
#> ||                                                                            ||
#> ||                Running time : 1.7 minutes                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> [1] "Index directory already exists remove it if you want to recreate it"
#> 
#>         ==========     _____ _    _ ____  _____  ______          _____  
#>         =====         / ____| |  | |  _ \|  __ \|  ____|   /\   |  __ \ 
#>           =====      | (___ | |  | | |_) | |__) | |__     /  \  | |  | |
#>             ====      \___ \| |  | |  _ <|  _  /|  __|   / /\ \ | |  | |
#>               ====    ____) | |__| | |_) | | \ \| |____ / ____ \| |__| |
#>         ==========   |_____/ \____/|____/|_|  \_\______/_/    \_\_____/
#>        Rsubread 2.20.0
#> 
#> //================================= setting ==================================\\
#> ||                                                                            ||
#> || Function      : Read alignment (DNA-Seq)                                   ||
#> || Input file    : prepro.PPP-BG.1M_CC_valid.fastq.gz                         ||
#> || Output file   : prepro.PPP-BG.1M_CC_valid.fastq.gz.bam (BAM), Sorted       ||
#> || Index name    : index                                                      ||
#> ||                                                                            ||
#> ||                    ------------------------------------                    ||
#> ||                                                                            ||
#> ||                               Threads : 1                                  ||
#> ||                          Phred offset : 33                                 ||
#> ||                             Min votes : 3 / 10                             ||
#> ||                        Max mismatches : 1                                  ||
#> ||                      Max indel length : 0                                  ||
#> ||            Report multi-mapping reads : no                                 ||
#> || Max alignments per multi-mapping read : 1                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> //================ Running (01-Nov-2025 17:39:02, pid=392792) ================\\
#> ||                                                                            ||
#> || Check the input reads.                                                     ||
#> || The input file contains base space reads.                                  ||
#> || Initialise the memory objects.                                             ||
#> || Estimate the mean read length.                                             ||
#> || WARNING  - The specified Phred score offset (33) seems incorrect.          ||
#> ||            ASCII values of the quality scores of read bases included in    ||
#> ||            the first 3000 reads were found to be within the range of       ||
#> ||            [63,63].                                                        ||
#> ||                                                                            ||
#> || Create the output BAM file.                                                ||
#> || Check the index.                                                           ||
#> || Init the voting space.                                                     ||
#> || Global environment is initialised.                                         ||
#> || Load the 1-th index block...                                               ||
#> || The index block has been loaded.                                           ||
#> || Start read mapping in chunk.                                               ||
#> ||    0% completed, 1.1 mins elapsed, rate=102.4k reads per second            ||
#> ||    6% completed, 1.2 mins elapsed, rate=113.6k reads per second            ||
#> ||   13% completed, 1.2 mins elapsed, rate=114.8k reads per second            ||
#> ||   20% completed, 1.2 mins elapsed, rate=115.3k reads per second            ||
#> ||   26% completed, 1.2 mins elapsed, rate=115.7k reads per second            ||
#> ||   33% completed, 1.2 mins elapsed, rate=115.7k reads per second            ||
#> ||   40% completed, 1.3 mins elapsed, rate=115.9k reads per second            ||
#> ||   46% completed, 1.3 mins elapsed, rate=116.0k reads per second            ||
#> ||   53% completed, 1.3 mins elapsed, rate=116.1k reads per second            ||
#> ||   60% completed, 1.3 mins elapsed, rate=115.5k reads per second            ||
#> ||   66% completed, 1.3 mins elapsed, rate=115.6k reads per second            ||
#> ||   70% completed, 1.4 mins elapsed, rate=17.1k reads per second             ||
#> ||   73% completed, 1.4 mins elapsed, rate=17.7k reads per second             ||
#> ||   76% completed, 1.4 mins elapsed, rate=18.3k reads per second             ||
#> ||   80% completed, 1.4 mins elapsed, rate=18.8k reads per second             ||
#> ||   83% completed, 1.4 mins elapsed, rate=19.4k reads per second             ||
#> ||   86% completed, 1.5 mins elapsed, rate=19.9k reads per second             ||
#> ||   90% completed, 1.5 mins elapsed, rate=20.4k reads per second             ||
#> ||   93% completed, 1.5 mins elapsed, rate=20.9k reads per second             ||
#> ||   96% completed, 1.5 mins elapsed, rate=21.4k reads per second             ||
#> ||                                                                            ||
#> ||                           Completed successfully.                          ||
#> ||                                                                            ||
#> \\====================================    ====================================//
#> 
#> //================================   Summary =================================\\
#> ||                                                                            ||
#> ||                 Total reads : 2,000,000                                    ||
#> ||                      Mapped : 375,407 (18.8%)                              ||
#> ||             Uniquely mapped : 375,407                                      ||
#> ||               Multi-mapping : 0                                            ||
#> ||                                                                            ||
#> ||                    Unmapped : 1,624,593                                    ||
#> ||                                                                            ||
#> ||                      Indels : 0                                            ||
#> ||                                                                            ||
#> ||                     WARNING : Phred offset (33) incorrect?                 ||
#> ||                                                                            ||
#> ||                Running time : 1.6 minutes                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> [1] "Index directory already exists remove it if you want to recreate it"
#> 
#>         ==========     _____ _    _ ____  _____  ______          _____  
#>         =====         / ____| |  | |  _ \|  __ \|  ____|   /\   |  __ \ 
#>           =====      | (___ | |  | | |_) | |__) | |__     /  \  | |  | |
#>             ====      \___ \| |  | |  _ <|  _  /|  __|   / /\ \ | |  | |
#>               ====    ____) | |__| | |_) | | \ \| |____ / ____ \| |__| |
#>         ==========   |_____/ \____/|____/|_|  \_\______/_/    \_\_____/
#>        Rsubread 2.20.0
#> 
#> //================================= setting ==================================\\
#> ||                                                                            ||
#> || Function      : Read alignment (DNA-Seq)                                   ||
#> || Input file    : prepro.PPP-BG.1M_CT_valid.fastq.gz                         ||
#> || Output file   : prepro.PPP-BG.1M_CT_valid.fastq.gz.bam (BAM), Sorted       ||
#> || Index name    : index                                                      ||
#> ||                                                                            ||
#> ||                    ------------------------------------                    ||
#> ||                                                                            ||
#> ||                               Threads : 1                                  ||
#> ||                          Phred offset : 33                                 ||
#> ||                             Min votes : 3 / 10                             ||
#> ||                        Max mismatches : 1                                  ||
#> ||                      Max indel length : 0                                  ||
#> ||            Report multi-mapping reads : no                                 ||
#> || Max alignments per multi-mapping read : 1                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> //================ Running (01-Nov-2025 17:40:37, pid=392792) ================\\
#> ||                                                                            ||
#> || Check the input reads.                                                     ||
#> || The input file contains base space reads.                                  ||
#> || Initialise the memory objects.                                             ||
#> || Estimate the mean read length.                                             ||
#> || WARNING  - The specified Phred score offset (33) seems incorrect.          ||
#> ||            ASCII values of the quality scores of read bases included in    ||
#> ||            the first 3000 reads were found to be within the range of       ||
#> ||            [63,63].                                                        ||
#> ||                                                                            ||
#> || Create the output BAM file.                                                ||
#> || Check the index.                                                           ||
#> || Init the voting space.                                                     ||
#> || Global environment is initialised.                                         ||
#> || Load the 1-th index block...                                               ||
#> || The index block has been loaded.                                           ||
#> || Start read mapping in chunk.                                               ||
#> ||    0% completed, 1.1 mins elapsed, rate=103.0k reads per second            ||
#> ||    6% completed, 1.2 mins elapsed, rate=115.3k reads per second            ||
#> ||   13% completed, 1.2 mins elapsed, rate=115.6k reads per second            ||
#> ||   20% completed, 1.2 mins elapsed, rate=115.9k reads per second            ||
#> ||   26% completed, 1.2 mins elapsed, rate=116.0k reads per second            ||
#> ||   33% completed, 1.2 mins elapsed, rate=115.9k reads per second            ||
#> ||   40% completed, 1.3 mins elapsed, rate=116.1k reads per second            ||
#> ||   46% completed, 1.3 mins elapsed, rate=116.0k reads per second            ||
#> ||   53% completed, 1.3 mins elapsed, rate=115.5k reads per second            ||
#> ||   60% completed, 1.3 mins elapsed, rate=115.5k reads per second            ||
#> ||   69% completed, 1.4 mins elapsed, rate=17.1k reads per second             ||
#> ||   73% completed, 1.4 mins elapsed, rate=17.7k reads per second             ||
#> ||   76% completed, 1.4 mins elapsed, rate=18.3k reads per second             ||
#> ||   80% completed, 1.4 mins elapsed, rate=18.8k reads per second             ||
#> ||   83% completed, 1.4 mins elapsed, rate=19.4k reads per second             ||
#> ||   86% completed, 1.5 mins elapsed, rate=19.9k reads per second             ||
#> ||   90% completed, 1.5 mins elapsed, rate=20.4k reads per second             ||
#> ||   93% completed, 1.5 mins elapsed, rate=20.9k reads per second             ||
#> ||   96% completed, 1.5 mins elapsed, rate=21.4k reads per second             ||
#> ||                                                                            ||
#> ||                           Completed successfully.                          ||
#> ||                                                                            ||
#> \\====================================    ====================================//
#> 
#> //================================   Summary =================================\\
#> ||                                                                            ||
#> ||                 Total reads : 2,000,000                                    ||
#> ||                      Mapped : 374,947 (18.7%)                              ||
#> ||             Uniquely mapped : 374,947                                      ||
#> ||               Multi-mapping : 0                                            ||
#> ||                                                                            ||
#> ||                    Unmapped : 1,625,053                                    ||
#> ||                                                                            ||
#> ||                      Indels : 0                                            ||
#> ||                                                                            ||
#> ||                     WARNING : Phred offset (33) incorrect?                 ||
#> ||                                                                            ||
#> ||                Running time : 1.6 minutes                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> [1] "Index directory already exists remove it if you want to recreate it"
#> 
#>         ==========     _____ _    _ ____  _____  ______          _____  
#>         =====         / ____| |  | |  _ \|  __ \|  ____|   /\   |  __ \ 
#>           =====      | (___ | |  | | |_) | |__) | |__     /  \  | |  | |
#>             ====      \___ \| |  | |  _ <|  _  /|  __|   / /\ \ | |  | |
#>               ====    ____) | |__| | |_) | | \ \| |____ / ____ \| |__| |
#>         ==========   |_____/ \____/|____/|_|  \_\______/_/    \_\_____/
#>        Rsubread 2.20.0
#> 
#> //================================= setting ==================================\\
#> ||                                                                            ||
#> || Function      : Read alignment (DNA-Seq)                                   ||
#> || Input file    : prepro.PPP-BG.5M_AA_valid.fastq.gz                         ||
#> || Output file   : prepro.PPP-BG.5M_AA_valid.fastq.gz.bam (BAM), Sorted       ||
#> || Index name    : index                                                      ||
#> ||                                                                            ||
#> ||                    ------------------------------------                    ||
#> ||                                                                            ||
#> ||                               Threads : 1                                  ||
#> ||                          Phred offset : 33                                 ||
#> ||                             Min votes : 3 / 10                             ||
#> ||                        Max mismatches : 1                                  ||
#> ||                      Max indel length : 0                                  ||
#> ||            Report multi-mapping reads : no                                 ||
#> || Max alignments per multi-mapping read : 1                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> //================ Running (01-Nov-2025 17:42:14, pid=392792) ================\\
#> ||                                                                            ||
#> || Check the input reads.                                                     ||
#> || The input file contains base space reads.                                  ||
#> || Initialise the memory objects.                                             ||
#> || Estimate the mean read length.                                             ||
#> || WARNING  - The specified Phred score offset (33) seems incorrect.          ||
#> ||            ASCII values of the quality scores of read bases included in    ||
#> ||            the first 3000 reads were found to be within the range of       ||
#> ||            [63,63].                                                        ||
#> ||                                                                            ||
#> || Create the output BAM file.                                                ||
#> || Check the index.                                                           ||
#> || Init the voting space.                                                     ||
#> || Global environment is initialised.                                         ||
#> || Load the 1-th index block...                                               ||
#> || The index block has been loaded.                                           ||
#> || Start read mapping in chunk.                                               ||
#> ||    0% completed, 1.1 mins elapsed, rate=98.2k reads per second             ||
#> ||    6% completed, 1.2 mins elapsed, rate=107.9k reads per second            ||
#> ||   12% completed, 1.3 mins elapsed, rate=107.5k reads per second            ||
#> ||   19% completed, 1.4 mins elapsed, rate=107.3k reads per second            ||
#> ||   26% completed, 1.6 mins elapsed, rate=107.2k reads per second            ||
#> ||   32% completed, 1.7 mins elapsed, rate=106.7k reads per second            ||
#> ||   39% completed, 1.8 mins elapsed, rate=105.9k reads per second            ||
#> ||   46% completed, 1.9 mins elapsed, rate=105.2k reads per second            ||
#> ||   48% completed, 1.9 mins elapsed, rate=41.9k reads per second             ||
#> ||   51% completed, 2.0 mins elapsed, rate=42.6k reads per second             ||
#> ||   53% completed, 2.1 mins elapsed, rate=43.3k reads per second             ||
#> ||   55% completed, 2.1 mins elapsed, rate=44.1k reads per second             ||
#> ||   58% completed, 2.2 mins elapsed, rate=44.7k reads per second             ||
#> ||   60% completed, 2.2 mins elapsed, rate=45.4k reads per second             ||
#> ||   62% completed, 2.3 mins elapsed, rate=45.9k reads per second             ||
#> ||   65% completed, 2.3 mins elapsed, rate=46.4k reads per second             ||
#> ||   67% completed, 2.4 mins elapsed, rate=46.8k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   69% completed, 3.2 mins elapsed, rate=56.9k reads per second             ||
#> ||   76% completed, 3.4 mins elapsed, rate=55.6k reads per second             ||
#> ||   83% completed, 3.6 mins elapsed, rate=57.1k reads per second             ||
#> ||   90% completed, 3.7 mins elapsed, rate=40.8k reads per second             ||
#> ||   91% completed, 3.7 mins elapsed, rate=41.0k reads per second             ||
#> ||   92% completed, 3.8 mins elapsed, rate=41.2k reads per second             ||
#> ||   93% completed, 3.8 mins elapsed, rate=41.4k reads per second             ||
#> ||   94% completed, 3.8 mins elapsed, rate=41.6k reads per second             ||
#> ||   95% completed, 3.8 mins elapsed, rate=41.8k reads per second             ||
#> ||   96% completed, 3.9 mins elapsed, rate=41.9k reads per second             ||
#> ||   97% completed, 3.9 mins elapsed, rate=42.1k reads per second             ||
#> ||   98% completed, 3.9 mins elapsed, rate=42.3k reads per second             ||
#> ||                                                                            ||
#> ||                           Completed successfully.                          ||
#> ||                                                                            ||
#> \\====================================    ====================================//
#> 
#> //================================   Summary =================================\\
#> ||                                                                            ||
#> ||                 Total reads : 10,000,000                                   ||
#> ||                      Mapped : 7,118,288 (71.2%)                            ||
#> ||             Uniquely mapped : 7,118,288                                    ||
#> ||               Multi-mapping : 0                                            ||
#> ||                                                                            ||
#> ||                    Unmapped : 2,881,712                                    ||
#> ||                                                                            ||
#> ||                      Indels : 0                                            ||
#> ||                                                                            ||
#> ||                     WARNING : Phred offset (33) incorrect?                 ||
#> ||                                                                            ||
#> ||                Running time : 4.2 minutes                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> [1] "Index directory already exists remove it if you want to recreate it"
#> 
#>         ==========     _____ _    _ ____  _____  ______          _____  
#>         =====         / ____| |  | |  _ \|  __ \|  ____|   /\   |  __ \ 
#>           =====      | (___ | |  | | |_) | |__) | |__     /  \  | |  | |
#>             ====      \___ \| |  | |  _ <|  _  /|  __|   / /\ \ | |  | |
#>               ====    ____) | |__| | |_) | | \ \| |____ / ____ \| |__| |
#>         ==========   |_____/ \____/|____/|_|  \_\______/_/    \_\_____/
#>        Rsubread 2.20.0
#> 
#> //================================= setting ==================================\\
#> ||                                                                            ||
#> || Function      : Read alignment (DNA-Seq)                                   ||
#> || Input file    : prepro.PPP-BG.5M_AC_valid.fastq.gz                         ||
#> || Output file   : prepro.PPP-BG.5M_AC_valid.fastq.gz.bam (BAM), Sorted       ||
#> || Index name    : index                                                      ||
#> ||                                                                            ||
#> ||                    ------------------------------------                    ||
#> ||                                                                            ||
#> ||                               Threads : 1                                  ||
#> ||                          Phred offset : 33                                 ||
#> ||                             Min votes : 3 / 10                             ||
#> ||                        Max mismatches : 1                                  ||
#> ||                      Max indel length : 0                                  ||
#> ||            Report multi-mapping reads : no                                 ||
#> || Max alignments per multi-mapping read : 1                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> //================ Running (01-Nov-2025 17:46:24, pid=392792) ================\\
#> ||                                                                            ||
#> || Check the input reads.                                                     ||
#> || The input file contains base space reads.                                  ||
#> || Initialise the memory objects.                                             ||
#> || Estimate the mean read length.                                             ||
#> || WARNING  - The specified Phred score offset (33) seems incorrect.          ||
#> ||            ASCII values of the quality scores of read bases included in    ||
#> ||            the first 3000 reads were found to be within the range of       ||
#> ||            [63,63].                                                        ||
#> ||                                                                            ||
#> || Create the output BAM file.                                                ||
#> || Check the index.                                                           ||
#> || Init the voting space.                                                     ||
#> || Global environment is initialised.                                         ||
#> || Load the 1-th index block...                                               ||
#> || The index block has been loaded.                                           ||
#> || Start read mapping in chunk.                                               ||
#> ||    0% completed, 1.1 mins elapsed, rate=101.1k reads per second            ||
#> ||    6% completed, 1.2 mins elapsed, rate=109.5k reads per second            ||
#> ||   12% completed, 1.3 mins elapsed, rate=109.3k reads per second            ||
#> ||   19% completed, 1.4 mins elapsed, rate=109.4k reads per second            ||
#> ||   26% completed, 1.5 mins elapsed, rate=108.3k reads per second            ||
#> ||   32% completed, 1.7 mins elapsed, rate=107.6k reads per second            ||
#> ||   39% completed, 1.8 mins elapsed, rate=107.8k reads per second            ||
#> ||   46% completed, 1.9 mins elapsed, rate=108.0k reads per second            ||
#> ||   48% completed, 1.9 mins elapsed, rate=42.5k reads per second             ||
#> ||   51% completed, 2.0 mins elapsed, rate=43.3k reads per second             ||
#> ||   53% completed, 2.0 mins elapsed, rate=44.1k reads per second             ||
#> ||   55% completed, 2.1 mins elapsed, rate=44.8k reads per second             ||
#> ||   58% completed, 2.1 mins elapsed, rate=45.6k reads per second             ||
#> ||   60% completed, 2.2 mins elapsed, rate=46.2k reads per second             ||
#> ||   62% completed, 2.2 mins elapsed, rate=46.9k reads per second             ||
#> ||   65% completed, 2.3 mins elapsed, rate=47.5k reads per second             ||
#> ||   67% completed, 2.3 mins elapsed, rate=48.1k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   69% completed, 2.6 mins elapsed, rate=79.6k reads per second             ||
#> ||   76% completed, 2.8 mins elapsed, rate=77.6k reads per second             ||
#> ||   83% completed, 2.9 mins elapsed, rate=78.8k reads per second             ||
#> ||   90% completed, 3.0 mins elapsed, rate=49.8k reads per second             ||
#> ||   91% completed, 3.1 mins elapsed, rate=50.0k reads per second             ||
#> ||   92% completed, 3.1 mins elapsed, rate=50.2k reads per second             ||
#> ||   93% completed, 3.1 mins elapsed, rate=50.4k reads per second             ||
#> ||   94% completed, 3.1 mins elapsed, rate=50.5k reads per second             ||
#> ||   95% completed, 3.2 mins elapsed, rate=50.6k reads per second             ||
#> ||   96% completed, 3.2 mins elapsed, rate=50.8k reads per second             ||
#> ||   97% completed, 3.2 mins elapsed, rate=51.0k reads per second             ||
#> ||   98% completed, 3.2 mins elapsed, rate=51.2k reads per second             ||
#> ||                                                                            ||
#> ||                           Completed successfully.                          ||
#> ||                                                                            ||
#> \\====================================    ====================================//
#> 
#> //================================   Summary =================================\\
#> ||                                                                            ||
#> ||                 Total reads : 10,000,000                                   ||
#> ||                      Mapped : 7,116,281 (71.2%)                            ||
#> ||             Uniquely mapped : 7,116,281                                    ||
#> ||               Multi-mapping : 0                                            ||
#> ||                                                                            ||
#> ||                    Unmapped : 2,883,719                                    ||
#> ||                                                                            ||
#> ||                      Indels : 0                                            ||
#> ||                                                                            ||
#> ||                     WARNING : Phred offset (33) incorrect?                 ||
#> ||                                                                            ||
#> ||                Running time : 3.5 minutes                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> [1] "Index directory already exists remove it if you want to recreate it"
#> 
#>         ==========     _____ _    _ ____  _____  ______          _____  
#>         =====         / ____| |  | |  _ \|  __ \|  ____|   /\   |  __ \ 
#>           =====      | (___ | |  | | |_) | |__) | |__     /  \  | |  | |
#>             ====      \___ \| |  | |  _ <|  _  /|  __|   / /\ \ | |  | |
#>               ====    ____) | |__| | |_) | | \ \| |____ / ____ \| |__| |
#>         ==========   |_____/ \____/|____/|_|  \_\______/_/    \_\_____/
#>        Rsubread 2.20.0
#> 
#> //================================= setting ==================================\\
#> ||                                                                            ||
#> || Function      : Read alignment (DNA-Seq)                                   ||
#> || Input file    : prepro.PPP-BG.5M_AT_valid.fastq.gz                         ||
#> || Output file   : prepro.PPP-BG.5M_AT_valid.fastq.gz.bam (BAM), Sorted       ||
#> || Index name    : index                                                      ||
#> ||                                                                            ||
#> ||                    ------------------------------------                    ||
#> ||                                                                            ||
#> ||                               Threads : 1                                  ||
#> ||                          Phred offset : 33                                 ||
#> ||                             Min votes : 3 / 10                             ||
#> ||                        Max mismatches : 1                                  ||
#> ||                      Max indel length : 0                                  ||
#> ||            Report multi-mapping reads : no                                 ||
#> || Max alignments per multi-mapping read : 1                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> //================ Running (01-Nov-2025 17:49:55, pid=392792) ================\\
#> ||                                                                            ||
#> || Check the input reads.                                                     ||
#> || The input file contains base space reads.                                  ||
#> || Initialise the memory objects.                                             ||
#> || Estimate the mean read length.                                             ||
#> || WARNING  - The specified Phred score offset (33) seems incorrect.          ||
#> ||            ASCII values of the quality scores of read bases included in    ||
#> ||            the first 3000 reads were found to be within the range of       ||
#> ||            [63,63].                                                        ||
#> ||                                                                            ||
#> || Create the output BAM file.                                                ||
#> || Check the index.                                                           ||
#> || Init the voting space.                                                     ||
#> || Global environment is initialised.                                         ||
#> || Load the 1-th index block...                                               ||
#> || The index block has been loaded.                                           ||
#> || Start read mapping in chunk.                                               ||
#> ||    0% completed, 1.1 mins elapsed, rate=99.9k reads per second             ||
#> ||    6% completed, 1.2 mins elapsed, rate=109.7k reads per second            ||
#> ||   12% completed, 1.3 mins elapsed, rate=109.4k reads per second            ||
#> ||   19% completed, 1.4 mins elapsed, rate=109.2k reads per second            ||
#> ||   26% completed, 1.5 mins elapsed, rate=109.2k reads per second            ||
#> ||   32% completed, 1.6 mins elapsed, rate=109.2k reads per second            ||
#> ||   39% completed, 1.7 mins elapsed, rate=109.3k reads per second            ||
#> ||   46% completed, 1.8 mins elapsed, rate=109.3k reads per second            ||
#> ||   48% completed, 1.9 mins elapsed, rate=42.7k reads per second             ||
#> ||   51% completed, 2.0 mins elapsed, rate=43.5k reads per second             ||
#> ||   53% completed, 2.0 mins elapsed, rate=44.2k reads per second             ||
#> ||   55% completed, 2.1 mins elapsed, rate=45.0k reads per second             ||
#> ||   58% completed, 2.1 mins elapsed, rate=45.7k reads per second             ||
#> ||   60% completed, 2.2 mins elapsed, rate=46.4k reads per second             ||
#> ||   62% completed, 2.2 mins elapsed, rate=47.0k reads per second             ||
#> ||   65% completed, 2.3 mins elapsed, rate=47.6k reads per second             ||
#> ||   67% completed, 2.3 mins elapsed, rate=48.2k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   69% completed, 2.5 mins elapsed, rate=85.3k reads per second             ||
#> ||   76% completed, 2.6 mins elapsed, rate=87.1k reads per second             ||
#> ||   83% completed, 2.7 mins elapsed, rate=88.7k reads per second             ||
#> ||   90% completed, 2.8 mins elapsed, rate=53.5k reads per second             ||
#> ||   91% completed, 2.9 mins elapsed, rate=53.7k reads per second             ||
#> ||   92% completed, 2.9 mins elapsed, rate=53.8k reads per second             ||
#> ||   93% completed, 2.9 mins elapsed, rate=54.0k reads per second             ||
#> ||   94% completed, 2.9 mins elapsed, rate=54.1k reads per second             ||
#> ||   95% completed, 2.9 mins elapsed, rate=54.3k reads per second             ||
#> ||   96% completed, 3.0 mins elapsed, rate=54.5k reads per second             ||
#> ||   97% completed, 3.0 mins elapsed, rate=54.6k reads per second             ||
#> ||   99% completed, 3.0 mins elapsed, rate=54.7k reads per second             ||
#> ||                                                                            ||
#> ||                           Completed successfully.                          ||
#> ||                                                                            ||
#> \\====================================    ====================================//
#> 
#> //================================   Summary =================================\\
#> ||                                                                            ||
#> ||                 Total reads : 10,000,000                                   ||
#> ||                      Mapped : 7,116,371 (71.2%)                            ||
#> ||             Uniquely mapped : 7,116,371                                    ||
#> ||               Multi-mapping : 0                                            ||
#> ||                                                                            ||
#> ||                    Unmapped : 2,883,629                                    ||
#> ||                                                                            ||
#> ||                      Indels : 0                                            ||
#> ||                                                                            ||
#> ||                     WARNING : Phred offset (33) incorrect?                 ||
#> ||                                                                            ||
#> ||                Running time : 3.3 minutes                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> [1] "Index directory already exists remove it if you want to recreate it"
#> 
#>         ==========     _____ _    _ ____  _____  ______          _____  
#>         =====         / ____| |  | |  _ \|  __ \|  ____|   /\   |  __ \ 
#>           =====      | (___ | |  | | |_) | |__) | |__     /  \  | |  | |
#>             ====      \___ \| |  | |  _ <|  _  /|  __|   / /\ \ | |  | |
#>               ====    ____) | |__| | |_) | | \ \| |____ / ____ \| |__| |
#>         ==========   |_____/ \____/|____/|_|  \_\______/_/    \_\_____/
#>        Rsubread 2.20.0
#> 
#> //================================= setting ==================================\\
#> ||                                                                            ||
#> || Function      : Read alignment (DNA-Seq)                                   ||
#> || Input file    : prepro.PPP-BG.5M_CA_valid.fastq.gz                         ||
#> || Output file   : prepro.PPP-BG.5M_CA_valid.fastq.gz.bam (BAM), Sorted       ||
#> || Index name    : index                                                      ||
#> ||                                                                            ||
#> ||                    ------------------------------------                    ||
#> ||                                                                            ||
#> ||                               Threads : 1                                  ||
#> ||                          Phred offset : 33                                 ||
#> ||                             Min votes : 3 / 10                             ||
#> ||                        Max mismatches : 1                                  ||
#> ||                      Max indel length : 0                                  ||
#> ||            Report multi-mapping reads : no                                 ||
#> || Max alignments per multi-mapping read : 1                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> //================ Running (01-Nov-2025 17:53:12, pid=392792) ================\\
#> ||                                                                            ||
#> || Check the input reads.                                                     ||
#> || The input file contains base space reads.                                  ||
#> || Initialise the memory objects.                                             ||
#> || Estimate the mean read length.                                             ||
#> || WARNING  - The specified Phred score offset (33) seems incorrect.          ||
#> ||            ASCII values of the quality scores of read bases included in    ||
#> ||            the first 3000 reads were found to be within the range of       ||
#> ||            [63,63].                                                        ||
#> ||                                                                            ||
#> || Create the output BAM file.                                                ||
#> || Check the index.                                                           ||
#> || Init the voting space.                                                     ||
#> || Global environment is initialised.                                         ||
#> || Load the 1-th index block...                                               ||
#> || The index block has been loaded.                                           ||
#> || Start read mapping in chunk.                                               ||
#> ||    0% completed, 1.2 mins elapsed, rate=103.8k reads per second            ||
#> ||    6% completed, 1.3 mins elapsed, rate=116.2k reads per second            ||
#> ||   13% completed, 1.4 mins elapsed, rate=116.4k reads per second            ||
#> ||   20% completed, 1.5 mins elapsed, rate=116.3k reads per second            ||
#> ||   26% completed, 1.6 mins elapsed, rate=116.3k reads per second            ||
#> ||   33% completed, 1.7 mins elapsed, rate=116.2k reads per second            ||
#> ||   40% completed, 1.8 mins elapsed, rate=116.1k reads per second            ||
#> ||   48% completed, 1.9 mins elapsed, rate=42.2k reads per second             ||
#> ||   51% completed, 2.0 mins elapsed, rate=42.8k reads per second             ||
#> ||   53% completed, 2.1 mins elapsed, rate=43.4k reads per second             ||
#> ||   55% completed, 2.1 mins elapsed, rate=44.0k reads per second             ||
#> ||   58% completed, 2.2 mins elapsed, rate=44.5k reads per second             ||
#> ||   60% completed, 2.2 mins elapsed, rate=45.0k reads per second             ||
#> ||   62% completed, 2.3 mins elapsed, rate=45.4k reads per second             ||
#> ||   65% completed, 2.4 mins elapsed, rate=45.9k reads per second             ||
#> ||   67% completed, 2.4 mins elapsed, rate=46.3k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   69% completed, 2.6 mins elapsed, rate=84.4k reads per second             ||
#> ||   76% completed, 2.7 mins elapsed, rate=86.4k reads per second             ||
#> ||   83% completed, 2.8 mins elapsed, rate=88.4k reads per second             ||
#> ||   90% completed, 2.9 mins elapsed, rate=52.5k reads per second             ||
#> ||   91% completed, 2.9 mins elapsed, rate=52.6k reads per second             ||
#> ||   92% completed, 2.9 mins elapsed, rate=52.7k reads per second             ||
#> ||   93% completed, 3.0 mins elapsed, rate=52.8k reads per second             ||
#> ||   94% completed, 3.0 mins elapsed, rate=52.8k reads per second             ||
#> ||   95% completed, 3.0 mins elapsed, rate=52.9k reads per second             ||
#> ||   96% completed, 3.0 mins elapsed, rate=53.0k reads per second             ||
#> ||   97% completed, 3.1 mins elapsed, rate=53.1k reads per second             ||
#> ||   98% completed, 3.1 mins elapsed, rate=53.2k reads per second             ||
#> ||                                                                            ||
#> ||                           Completed successfully.                          ||
#> ||                                                                            ||
#> \\====================================    ====================================//
#> 
#> //================================   Summary =================================\\
#> ||                                                                            ||
#> ||                 Total reads : 10,000,000                                   ||
#> ||                      Mapped : 1,873,121 (18.7%)                            ||
#> ||             Uniquely mapped : 1,873,121                                    ||
#> ||               Multi-mapping : 0                                            ||
#> ||                                                                            ||
#> ||                    Unmapped : 8,126,879                                    ||
#> ||                                                                            ||
#> ||                      Indels : 0                                            ||
#> ||                                                                            ||
#> ||                     WARNING : Phred offset (33) incorrect?                 ||
#> ||                                                                            ||
#> ||                Running time : 3.4 minutes                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> [1] "Index directory already exists remove it if you want to recreate it"
#> 
#>         ==========     _____ _    _ ____  _____  ______          _____  
#>         =====         / ____| |  | |  _ \|  __ \|  ____|   /\   |  __ \ 
#>           =====      | (___ | |  | | |_) | |__) | |__     /  \  | |  | |
#>             ====      \___ \| |  | |  _ <|  _  /|  __|   / /\ \ | |  | |
#>               ====    ____) | |__| | |_) | | \ \| |____ / ____ \| |__| |
#>         ==========   |_____/ \____/|____/|_|  \_\______/_/    \_\_____/
#>        Rsubread 2.20.0
#> 
#> //================================= setting ==================================\\
#> ||                                                                            ||
#> || Function      : Read alignment (DNA-Seq)                                   ||
#> || Input file    : prepro.PPP-BG.5M_CC_valid.fastq.gz                         ||
#> || Output file   : prepro.PPP-BG.5M_CC_valid.fastq.gz.bam (BAM), Sorted       ||
#> || Index name    : index                                                      ||
#> ||                                                                            ||
#> ||                    ------------------------------------                    ||
#> ||                                                                            ||
#> ||                               Threads : 1                                  ||
#> ||                          Phred offset : 33                                 ||
#> ||                             Min votes : 3 / 10                             ||
#> ||                        Max mismatches : 1                                  ||
#> ||                      Max indel length : 0                                  ||
#> ||            Report multi-mapping reads : no                                 ||
#> || Max alignments per multi-mapping read : 1                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> //================ Running (01-Nov-2025 17:56:33, pid=392792) ================\\
#> ||                                                                            ||
#> || Check the input reads.                                                     ||
#> || The input file contains base space reads.                                  ||
#> || Initialise the memory objects.                                             ||
#> || Estimate the mean read length.                                             ||
#> || WARNING  - The specified Phred score offset (33) seems incorrect.          ||
#> ||            ASCII values of the quality scores of read bases included in    ||
#> ||            the first 3000 reads were found to be within the range of       ||
#> ||            [63,63].                                                        ||
#> ||                                                                            ||
#> || Create the output BAM file.                                                ||
#> || Check the index.                                                           ||
#> || Init the voting space.                                                     ||
#> || Global environment is initialised.                                         ||
#> || Load the 1-th index block...                                               ||
#> || The index block has been loaded.                                           ||
#> || Start read mapping in chunk.                                               ||
#> ||    0% completed, 1.1 mins elapsed, rate=103.0k reads per second            ||
#> ||    6% completed, 1.2 mins elapsed, rate=115.5k reads per second            ||
#> ||   13% completed, 1.3 mins elapsed, rate=115.5k reads per second            ||
#> ||   19% completed, 1.4 mins elapsed, rate=114.3k reads per second            ||
#> ||   26% completed, 1.5 mins elapsed, rate=114.6k reads per second            ||
#> ||   33% completed, 1.6 mins elapsed, rate=114.8k reads per second            ||
#> ||   39% completed, 1.7 mins elapsed, rate=115.0k reads per second            ||
#> ||   46% completed, 1.8 mins elapsed, rate=114.9k reads per second            ||
#> ||   48% completed, 1.9 mins elapsed, rate=43.2k reads per second             ||
#> ||   51% completed, 2.0 mins elapsed, rate=43.8k reads per second             ||
#> ||   53% completed, 2.0 mins elapsed, rate=44.3k reads per second             ||
#> ||   55% completed, 2.1 mins elapsed, rate=44.9k reads per second             ||
#> ||   58% completed, 2.1 mins elapsed, rate=45.4k reads per second             ||
#> ||   60% completed, 2.2 mins elapsed, rate=45.9k reads per second             ||
#> ||   62% completed, 2.3 mins elapsed, rate=46.3k reads per second             ||
#> ||   65% completed, 2.3 mins elapsed, rate=46.7k reads per second             ||
#> ||   67% completed, 2.4 mins elapsed, rate=47.1k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   69% completed, 2.5 mins elapsed, rate=84.4k reads per second             ||
#> ||   76% completed, 2.6 mins elapsed, rate=86.5k reads per second             ||
#> ||   83% completed, 2.7 mins elapsed, rate=88.4k reads per second             ||
#> ||   90% completed, 2.8 mins elapsed, rate=53.4k reads per second             ||
#> ||   91% completed, 2.9 mins elapsed, rate=53.5k reads per second             ||
#> ||   92% completed, 2.9 mins elapsed, rate=53.5k reads per second             ||
#> ||   93% completed, 2.9 mins elapsed, rate=53.6k reads per second             ||
#> ||   94% completed, 2.9 mins elapsed, rate=53.7k reads per second             ||
#> ||   95% completed, 3.0 mins elapsed, rate=53.8k reads per second             ||
#> ||   96% completed, 3.0 mins elapsed, rate=53.9k reads per second             ||
#> ||   97% completed, 3.0 mins elapsed, rate=53.9k reads per second             ||
#> ||   99% completed, 3.1 mins elapsed, rate=54.0k reads per second             ||
#> ||                                                                            ||
#> ||                           Completed successfully.                          ||
#> ||                                                                            ||
#> \\====================================    ====================================//
#> 
#> //================================   Summary =================================\\
#> ||                                                                            ||
#> ||                 Total reads : 10,000,000                                   ||
#> ||                      Mapped : 1,876,176 (18.8%)                            ||
#> ||             Uniquely mapped : 1,876,176                                    ||
#> ||               Multi-mapping : 0                                            ||
#> ||                                                                            ||
#> ||                    Unmapped : 8,123,824                                    ||
#> ||                                                                            ||
#> ||                      Indels : 0                                            ||
#> ||                                                                            ||
#> ||                     WARNING : Phred offset (33) incorrect?                 ||
#> ||                                                                            ||
#> ||                Running time : 3.3 minutes                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> [1] "Index directory already exists remove it if you want to recreate it"
#> 
#>         ==========     _____ _    _ ____  _____  ______          _____  
#>         =====         / ____| |  | |  _ \|  __ \|  ____|   /\   |  __ \ 
#>           =====      | (___ | |  | | |_) | |__) | |__     /  \  | |  | |
#>             ====      \___ \| |  | |  _ <|  _  /|  __|   / /\ \ | |  | |
#>               ====    ____) | |__| | |_) | | \ \| |____ / ____ \| |__| |
#>         ==========   |_____/ \____/|____/|_|  \_\______/_/    \_\_____/
#>        Rsubread 2.20.0
#> 
#> //================================= setting ==================================\\
#> ||                                                                            ||
#> || Function      : Read alignment (DNA-Seq)                                   ||
#> || Input file    : prepro.PPP-BG.5M_CT_valid.fastq.gz                         ||
#> || Output file   : prepro.PPP-BG.5M_CT_valid.fastq.gz.bam (BAM), Sorted       ||
#> || Index name    : index                                                      ||
#> ||                                                                            ||
#> ||                    ------------------------------------                    ||
#> ||                                                                            ||
#> ||                               Threads : 1                                  ||
#> ||                          Phred offset : 33                                 ||
#> ||                             Min votes : 3 / 10                             ||
#> ||                        Max mismatches : 1                                  ||
#> ||                      Max indel length : 0                                  ||
#> ||            Report multi-mapping reads : no                                 ||
#> || Max alignments per multi-mapping read : 1                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> //================ Running (01-Nov-2025 17:59:52, pid=392792) ================\\
#> ||                                                                            ||
#> || Check the input reads.                                                     ||
#> || The input file contains base space reads.                                  ||
#> || Initialise the memory objects.                                             ||
#> || Estimate the mean read length.                                             ||
#> || WARNING  - The specified Phred score offset (33) seems incorrect.          ||
#> ||            ASCII values of the quality scores of read bases included in    ||
#> ||            the first 3000 reads were found to be within the range of       ||
#> ||            [63,63].                                                        ||
#> ||                                                                            ||
#> || Create the output BAM file.                                                ||
#> || Check the index.                                                           ||
#> || Init the voting space.                                                     ||
#> || Global environment is initialised.                                         ||
#> || Load the 1-th index block...                                               ||
#> || The index block has been loaded.                                           ||
#> || Start read mapping in chunk.                                               ||
#> ||    0% completed, 1.1 mins elapsed, rate=104.0k reads per second            ||
#> ||    6% completed, 1.2 mins elapsed, rate=116.0k reads per second            ||
#> ||   13% completed, 1.3 mins elapsed, rate=115.9k reads per second            ||
#> ||   20% completed, 1.4 mins elapsed, rate=115.8k reads per second            ||
#> ||   26% completed, 1.5 mins elapsed, rate=115.8k reads per second            ||
#> ||   33% completed, 1.6 mins elapsed, rate=115.7k reads per second            ||
#> ||   40% completed, 1.7 mins elapsed, rate=115.8k reads per second            ||
#> ||   48% completed, 1.9 mins elapsed, rate=43.4k reads per second             ||
#> ||   51% completed, 1.9 mins elapsed, rate=43.9k reads per second             ||
#> ||   53% completed, 2.0 mins elapsed, rate=44.5k reads per second             ||
#> ||   55% completed, 2.1 mins elapsed, rate=45.1k reads per second             ||
#> ||   58% completed, 2.1 mins elapsed, rate=45.6k reads per second             ||
#> ||   60% completed, 2.2 mins elapsed, rate=46.0k reads per second             ||
#> ||   62% completed, 2.3 mins elapsed, rate=46.5k reads per second             ||
#> ||   65% completed, 2.3 mins elapsed, rate=46.9k reads per second             ||
#> ||   67% completed, 2.4 mins elapsed, rate=47.3k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   69% completed, 2.5 mins elapsed, rate=84.9k reads per second             ||
#> ||   76% completed, 2.6 mins elapsed, rate=87.0k reads per second             ||
#> ||   83% completed, 2.7 mins elapsed, rate=88.9k reads per second             ||
#> ||   90% completed, 2.8 mins elapsed, rate=53.6k reads per second             ||
#> ||   91% completed, 2.9 mins elapsed, rate=53.7k reads per second             ||
#> ||   92% completed, 2.9 mins elapsed, rate=53.8k reads per second             ||
#> ||   93% completed, 2.9 mins elapsed, rate=53.8k reads per second             ||
#> ||   94% completed, 2.9 mins elapsed, rate=53.9k reads per second             ||
#> ||   95% completed, 3.0 mins elapsed, rate=54.0k reads per second             ||
#> ||   96% completed, 3.0 mins elapsed, rate=54.1k reads per second             ||
#> ||   97% completed, 3.0 mins elapsed, rate=54.2k reads per second             ||
#> ||   98% completed, 3.0 mins elapsed, rate=54.2k reads per second             ||
#> ||                                                                            ||
#> ||                           Completed successfully.                          ||
#> ||                                                                            ||
#> \\====================================    ====================================//
#> 
#> //================================   Summary =================================\\
#> ||                                                                            ||
#> ||                 Total reads : 10,000,000                                   ||
#> ||                      Mapped : 1,876,608 (18.8%)                            ||
#> ||             Uniquely mapped : 1,876,608                                    ||
#> ||               Multi-mapping : 0                                            ||
#> ||                                                                            ||
#> ||                    Unmapped : 8,123,392                                    ||
#> ||                                                                            ||
#> ||                      Indels : 0                                            ||
#> ||                                                                            ||
#> ||                     WARNING : Phred offset (33) incorrect?                 ||
#> ||                                                                            ||
#> ||                Running time : 3.3 minutes                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> [1] "Index directory already exists remove it if you want to recreate it"
#> 
#>         ==========     _____ _    _ ____  _____  ______          _____  
#>         =====         / ____| |  | |  _ \|  __ \|  ____|   /\   |  __ \ 
#>           =====      | (___ | |  | | |_) | |__) | |__     /  \  | |  | |
#>             ====      \___ \| |  | |  _ <|  _  /|  __|   / /\ \ | |  | |
#>               ====    ____) | |__| | |_) | | \ \| |____ / ____ \| |__| |
#>         ==========   |_____/ \____/|____/|_|  \_\______/_/    \_\_____/
#>        Rsubread 2.20.0
#> 
#> //================================= setting ==================================\\
#> ||                                                                            ||
#> || Function      : Read alignment (DNA-Seq)                                   ||
#> || Input file    : prepro.PPP-BG.10M_AA_valid.fastq.gz                        ||
#> || Output file   : prepro.PPP-BG.10M_AA_valid.fastq.gz.bam (BAM), Sorted      ||
#> || Index name    : index                                                      ||
#> ||                                                                            ||
#> ||                    ------------------------------------                    ||
#> ||                                                                            ||
#> ||                               Threads : 1                                  ||
#> ||                          Phred offset : 33                                 ||
#> ||                             Min votes : 3 / 10                             ||
#> ||                        Max mismatches : 1                                  ||
#> ||                      Max indel length : 0                                  ||
#> ||            Report multi-mapping reads : no                                 ||
#> || Max alignments per multi-mapping read : 1                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> //================ Running (01-Nov-2025 18:03:25, pid=392792) ================\\
#> ||                                                                            ||
#> || Check the input reads.                                                     ||
#> || The input file contains base space reads.                                  ||
#> || Initialise the memory objects.                                             ||
#> || Estimate the mean read length.                                             ||
#> || WARNING  - The specified Phred score offset (33) seems incorrect.          ||
#> ||            ASCII values of the quality scores of read bases included in    ||
#> ||            the first 3000 reads were found to be within the range of       ||
#> ||            [63,63].                                                        ||
#> ||                                                                            ||
#> || Create the output BAM file.                                                ||
#> || Check the index.                                                           ||
#> || Init the voting space.                                                     ||
#> || Global environment is initialised.                                         ||
#> || Load the 1-th index block...                                               ||
#> || The index block has been loaded.                                           ||
#> || Start read mapping in chunk.                                               ||
#> ||    0% completed, 1.1 mins elapsed, rate=96.2k reads per second             ||
#> ||    6% completed, 1.3 mins elapsed, rate=109.1k reads per second            ||
#> ||   12% completed, 1.5 mins elapsed, rate=109.2k reads per second            ||
#> ||   19% completed, 1.7 mins elapsed, rate=109.1k reads per second            ||
#> ||   24% completed, 1.9 mins elapsed, rate=42.6k reads per second             ||
#> ||   25% completed, 2.0 mins elapsed, rate=43.4k reads per second             ||
#> ||   26% completed, 2.0 mins elapsed, rate=44.2k reads per second             ||
#> ||   27% completed, 2.1 mins elapsed, rate=44.9k reads per second             ||
#> ||   29% completed, 2.1 mins elapsed, rate=45.6k reads per second             ||
#> ||   30% completed, 2.2 mins elapsed, rate=46.3k reads per second             ||
#> ||   31% completed, 2.2 mins elapsed, rate=47.0k reads per second             ||
#> ||   32% completed, 2.3 mins elapsed, rate=47.6k reads per second             ||
#> ||   33% completed, 2.3 mins elapsed, rate=48.2k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   34% completed, 2.5 mins elapsed, rate=84.4k reads per second             ||
#> ||   41% completed, 2.7 mins elapsed, rate=87.8k reads per second             ||
#> ||   48% completed, 2.9 mins elapsed, rate=90.6k reads per second             ||
#> ||   54% completed, 3.1 mins elapsed, rate=92.8k reads per second             ||
#> ||   59% completed, 3.3 mins elapsed, rate=60.6k reads per second             ||
#> ||   60% completed, 3.3 mins elapsed, rate=60.8k reads per second             ||
#> ||   61% completed, 3.4 mins elapsed, rate=61.0k reads per second             ||
#> ||   62% completed, 3.4 mins elapsed, rate=61.2k reads per second             ||
#> ||   64% completed, 3.5 mins elapsed, rate=61.4k reads per second             ||
#> ||   65% completed, 3.5 mins elapsed, rate=61.6k reads per second             ||
#> ||   66% completed, 3.6 mins elapsed, rate=61.8k reads per second             ||
#> ||   67% completed, 3.6 mins elapsed, rate=62.0k reads per second             ||
#> ||   68% completed, 3.7 mins elapsed, rate=62.2k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   69% completed, 3.9 mins elapsed, rate=85.9k reads per second             ||
#> ||   76% completed, 4.1 mins elapsed, rate=87.7k reads per second             ||
#> ||   83% completed, 4.3 mins elapsed, rate=89.3k reads per second             ||
#> ||   89% completed, 4.5 mins elapsed, rate=90.7k reads per second             ||
#> ||   90% completed, 4.5 mins elapsed, rate=67.4k reads per second             ||
#> ||   91% completed, 4.5 mins elapsed, rate=67.4k reads per second             ||
#> ||   92% completed, 4.6 mins elapsed, rate=67.5k reads per second             ||
#> ||   93% completed, 4.6 mins elapsed, rate=67.6k reads per second             ||
#> ||   94% completed, 4.7 mins elapsed, rate=67.7k reads per second             ||
#> ||   95% completed, 4.7 mins elapsed, rate=67.7k reads per second             ||
#> ||   96% completed, 4.8 mins elapsed, rate=67.8k reads per second             ||
#> ||   97% completed, 4.8 mins elapsed, rate=67.9k reads per second             ||
#> ||   98% completed, 4.9 mins elapsed, rate=67.9k reads per second             ||
#> ||                                                                            ||
#> ||                           Completed successfully.                          ||
#> ||                                                                            ||
#> \\====================================    ====================================//
#> 
#> //================================   Summary =================================\\
#> ||                                                                            ||
#> ||                 Total reads : 20,000,000                                   ||
#> ||                      Mapped : 14,236,795 (71.2%)                           ||
#> ||             Uniquely mapped : 14,236,795                                   ||
#> ||               Multi-mapping : 0                                            ||
#> ||                                                                            ||
#> ||                    Unmapped : 5,763,205                                    ||
#> ||                                                                            ||
#> ||                      Indels : 0                                            ||
#> ||                                                                            ||
#> ||                     WARNING : Phred offset (33) incorrect?                 ||
#> ||                                                                            ||
#> ||                Running time : 5.4 minutes                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> [1] "Index directory already exists remove it if you want to recreate it"
#> 
#>         ==========     _____ _    _ ____  _____  ______          _____  
#>         =====         / ____| |  | |  _ \|  __ \|  ____|   /\   |  __ \ 
#>           =====      | (___ | |  | | |_) | |__) | |__     /  \  | |  | |
#>             ====      \___ \| |  | |  _ <|  _  /|  __|   / /\ \ | |  | |
#>               ====    ____) | |__| | |_) | | \ \| |____ / ____ \| |__| |
#>         ==========   |_____/ \____/|____/|_|  \_\______/_/    \_\_____/
#>        Rsubread 2.20.0
#> 
#> //================================= setting ==================================\\
#> ||                                                                            ||
#> || Function      : Read alignment (DNA-Seq)                                   ||
#> || Input file    : prepro.PPP-BG.10M_AC_valid.fastq.gz                        ||
#> || Output file   : prepro.PPP-BG.10M_AC_valid.fastq.gz.bam (BAM), Sorted      ||
#> || Index name    : index                                                      ||
#> ||                                                                            ||
#> ||                    ------------------------------------                    ||
#> ||                                                                            ||
#> ||                               Threads : 1                                  ||
#> ||                          Phred offset : 33                                 ||
#> ||                             Min votes : 3 / 10                             ||
#> ||                        Max mismatches : 1                                  ||
#> ||                      Max indel length : 0                                  ||
#> ||            Report multi-mapping reads : no                                 ||
#> || Max alignments per multi-mapping read : 1                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> //================ Running (01-Nov-2025 18:08:51, pid=392792) ================\\
#> ||                                                                            ||
#> || Check the input reads.                                                     ||
#> || The input file contains base space reads.                                  ||
#> || Initialise the memory objects.                                             ||
#> || Estimate the mean read length.                                             ||
#> || WARNING  - The specified Phred score offset (33) seems incorrect.          ||
#> ||            ASCII values of the quality scores of read bases included in    ||
#> ||            the first 3000 reads were found to be within the range of       ||
#> ||            [63,63].                                                        ||
#> ||                                                                            ||
#> || Create the output BAM file.                                                ||
#> || Check the index.                                                           ||
#> || Init the voting space.                                                     ||
#> || Global environment is initialised.                                         ||
#> || Load the 1-th index block...                                               ||
#> || The index block has been loaded.                                           ||
#> || Start read mapping in chunk.                                               ||
#> ||    0% completed, 1.2 mins elapsed, rate=100.2k reads per second            ||
#> ||    6% completed, 1.3 mins elapsed, rate=109.1k reads per second            ||
#> ||   12% completed, 1.5 mins elapsed, rate=109.2k reads per second            ||
#> ||   19% completed, 1.7 mins elapsed, rate=109.2k reads per second            ||
#> ||   24% completed, 1.9 mins elapsed, rate=42.6k reads per second             ||
#> ||   25% completed, 2.0 mins elapsed, rate=43.4k reads per second             ||
#> ||   26% completed, 2.0 mins elapsed, rate=44.2k reads per second             ||
#> ||   27% completed, 2.1 mins elapsed, rate=44.9k reads per second             ||
#> ||   29% completed, 2.1 mins elapsed, rate=45.6k reads per second             ||
#> ||   30% completed, 2.2 mins elapsed, rate=46.3k reads per second             ||
#> ||   31% completed, 2.2 mins elapsed, rate=46.9k reads per second             ||
#> ||   32% completed, 2.3 mins elapsed, rate=47.5k reads per second             ||
#> ||   33% completed, 2.3 mins elapsed, rate=48.1k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   34% completed, 2.5 mins elapsed, rate=85.5k reads per second             ||
#> ||   41% completed, 2.7 mins elapsed, rate=88.9k reads per second             ||
#> ||   48% completed, 2.9 mins elapsed, rate=91.5k reads per second             ||
#> ||   54% completed, 3.1 mins elapsed, rate=93.7k reads per second             ||
#> ||   59% completed, 3.3 mins elapsed, rate=60.8k reads per second             ||
#> ||   60% completed, 3.3 mins elapsed, rate=61.0k reads per second             ||
#> ||   61% completed, 3.4 mins elapsed, rate=61.2k reads per second             ||
#> ||   62% completed, 3.4 mins elapsed, rate=61.4k reads per second             ||
#> ||   63% completed, 3.5 mins elapsed, rate=61.7k reads per second             ||
#> ||   65% completed, 3.5 mins elapsed, rate=61.9k reads per second             ||
#> ||   66% completed, 3.6 mins elapsed, rate=62.1k reads per second             ||
#> ||   67% completed, 3.6 mins elapsed, rate=62.2k reads per second             ||
#> ||   68% completed, 3.7 mins elapsed, rate=62.4k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   69% completed, 3.8 mins elapsed, rate=86.4k reads per second             ||
#> ||   76% completed, 4.0 mins elapsed, rate=88.2k reads per second             ||
#> ||   83% completed, 4.2 mins elapsed, rate=89.7k reads per second             ||
#> ||   89% completed, 4.4 mins elapsed, rate=90.9k reads per second             ||
#> ||   90% completed, 4.5 mins elapsed, rate=67.4k reads per second             ||
#> ||   91% completed, 4.5 mins elapsed, rate=67.5k reads per second             ||
#> ||   92% completed, 4.6 mins elapsed, rate=67.6k reads per second             ||
#> ||   93% completed, 4.6 mins elapsed, rate=67.6k reads per second             ||
#> ||   94% completed, 4.7 mins elapsed, rate=67.7k reads per second             ||
#> ||   95% completed, 4.7 mins elapsed, rate=67.8k reads per second             ||
#> ||   96% completed, 4.8 mins elapsed, rate=67.9k reads per second             ||
#> ||   97% completed, 4.8 mins elapsed, rate=67.9k reads per second             ||
#> ||   98% completed, 4.9 mins elapsed, rate=68.0k reads per second             ||
#> ||                                                                            ||
#> ||                           Completed successfully.                          ||
#> ||                                                                            ||
#> \\====================================    ====================================//
#> 
#> //================================   Summary =================================\\
#> ||                                                                            ||
#> ||                 Total reads : 20,000,000                                   ||
#> ||                      Mapped : 14,233,458 (71.2%)                           ||
#> ||             Uniquely mapped : 14,233,458                                   ||
#> ||               Multi-mapping : 0                                            ||
#> ||                                                                            ||
#> ||                    Unmapped : 5,766,542                                    ||
#> ||                                                                            ||
#> ||                      Indels : 0                                            ||
#> ||                                                                            ||
#> ||                     WARNING : Phred offset (33) incorrect?                 ||
#> ||                                                                            ||
#> ||                Running time : 5.4 minutes                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> [1] "Index directory already exists remove it if you want to recreate it"
#> 
#>         ==========     _____ _    _ ____  _____  ______          _____  
#>         =====         / ____| |  | |  _ \|  __ \|  ____|   /\   |  __ \ 
#>           =====      | (___ | |  | | |_) | |__) | |__     /  \  | |  | |
#>             ====      \___ \| |  | |  _ <|  _  /|  __|   / /\ \ | |  | |
#>               ====    ____) | |__| | |_) | | \ \| |____ / ____ \| |__| |
#>         ==========   |_____/ \____/|____/|_|  \_\______/_/    \_\_____/
#>        Rsubread 2.20.0
#> 
#> //================================= setting ==================================\\
#> ||                                                                            ||
#> || Function      : Read alignment (DNA-Seq)                                   ||
#> || Input file    : prepro.PPP-BG.10M_AT_valid.fastq.gz                        ||
#> || Output file   : prepro.PPP-BG.10M_AT_valid.fastq.gz.bam (BAM), Sorted      ||
#> || Index name    : index                                                      ||
#> ||                                                                            ||
#> ||                    ------------------------------------                    ||
#> ||                                                                            ||
#> ||                               Threads : 1                                  ||
#> ||                          Phred offset : 33                                 ||
#> ||                             Min votes : 3 / 10                             ||
#> ||                        Max mismatches : 1                                  ||
#> ||                      Max indel length : 0                                  ||
#> ||            Report multi-mapping reads : no                                 ||
#> || Max alignments per multi-mapping read : 1                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> //================ Running (01-Nov-2025 18:14:16, pid=392792) ================\\
#> ||                                                                            ||
#> || Check the input reads.                                                     ||
#> || The input file contains base space reads.                                  ||
#> || Initialise the memory objects.                                             ||
#> || Estimate the mean read length.                                             ||
#> || WARNING  - The specified Phred score offset (33) seems incorrect.          ||
#> ||            ASCII values of the quality scores of read bases included in    ||
#> ||            the first 3000 reads were found to be within the range of       ||
#> ||            [63,63].                                                        ||
#> ||                                                                            ||
#> || Create the output BAM file.                                                ||
#> || Check the index.                                                           ||
#> || Init the voting space.                                                     ||
#> || Global environment is initialised.                                         ||
#> || Load the 1-th index block...                                               ||
#> || The index block has been loaded.                                           ||
#> || Start read mapping in chunk.                                               ||
#> ||    0% completed, 1.1 mins elapsed, rate=100.2k reads per second            ||
#> ||    6% completed, 1.3 mins elapsed, rate=109.2k reads per second            ||
#> ||   12% completed, 1.5 mins elapsed, rate=109.2k reads per second            ||
#> ||   19% completed, 1.7 mins elapsed, rate=109.1k reads per second            ||
#> ||   24% completed, 1.9 mins elapsed, rate=42.6k reads per second             ||
#> ||   25% completed, 2.0 mins elapsed, rate=43.4k reads per second             ||
#> ||   26% completed, 2.0 mins elapsed, rate=44.2k reads per second             ||
#> ||   27% completed, 2.1 mins elapsed, rate=44.9k reads per second             ||
#> ||   29% completed, 2.1 mins elapsed, rate=45.7k reads per second             ||
#> ||   30% completed, 2.2 mins elapsed, rate=46.3k reads per second             ||
#> ||   31% completed, 2.2 mins elapsed, rate=47.0k reads per second             ||
#> ||   32% completed, 2.3 mins elapsed, rate=47.6k reads per second             ||
#> ||   33% completed, 2.3 mins elapsed, rate=48.2k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   34% completed, 2.5 mins elapsed, rate=84.4k reads per second             ||
#> ||   41% completed, 2.7 mins elapsed, rate=87.9k reads per second             ||
#> ||   48% completed, 2.9 mins elapsed, rate=90.7k reads per second             ||
#> ||   54% completed, 3.1 mins elapsed, rate=92.9k reads per second             ||
#> ||   59% completed, 3.3 mins elapsed, rate=60.5k reads per second             ||
#> ||   60% completed, 3.3 mins elapsed, rate=60.8k reads per second             ||
#> ||   61% completed, 3.4 mins elapsed, rate=61.0k reads per second             ||
#> ||   62% completed, 3.4 mins elapsed, rate=61.2k reads per second             ||
#> ||   63% completed, 3.5 mins elapsed, rate=61.4k reads per second             ||
#> ||   65% completed, 3.5 mins elapsed, rate=61.6k reads per second             ||
#> ||   66% completed, 3.6 mins elapsed, rate=61.8k reads per second             ||
#> ||   67% completed, 3.6 mins elapsed, rate=62.0k reads per second             ||
#> ||   68% completed, 3.7 mins elapsed, rate=62.2k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   69% completed, 3.9 mins elapsed, rate=86.0k reads per second             ||
#> ||   76% completed, 4.1 mins elapsed, rate=87.8k reads per second             ||
#> ||   83% completed, 4.3 mins elapsed, rate=89.4k reads per second             ||
#> ||   89% completed, 4.4 mins elapsed, rate=90.8k reads per second             ||
#> ||   90% completed, 4.5 mins elapsed, rate=67.4k reads per second             ||
#> ||   91% completed, 4.5 mins elapsed, rate=67.5k reads per second             ||
#> ||   92% completed, 4.6 mins elapsed, rate=67.6k reads per second             ||
#> ||   93% completed, 4.6 mins elapsed, rate=67.6k reads per second             ||
#> ||   94% completed, 4.7 mins elapsed, rate=67.7k reads per second             ||
#> ||   95% completed, 4.7 mins elapsed, rate=67.8k reads per second             ||
#> ||   96% completed, 4.8 mins elapsed, rate=67.8k reads per second             ||
#> ||   97% completed, 4.8 mins elapsed, rate=67.9k reads per second             ||
#> ||   98% completed, 4.9 mins elapsed, rate=68.0k reads per second             ||
#> ||                                                                            ||
#> ||                           Completed successfully.                          ||
#> ||                                                                            ||
#> \\====================================    ====================================//
#> 
#> //================================   Summary =================================\\
#> ||                                                                            ||
#> ||                 Total reads : 20,000,000                                   ||
#> ||                      Mapped : 14,234,627 (71.2%)                           ||
#> ||             Uniquely mapped : 14,234,627                                   ||
#> ||               Multi-mapping : 0                                            ||
#> ||                                                                            ||
#> ||                    Unmapped : 5,765,373                                    ||
#> ||                                                                            ||
#> ||                      Indels : 0                                            ||
#> ||                                                                            ||
#> ||                     WARNING : Phred offset (33) incorrect?                 ||
#> ||                                                                            ||
#> ||                Running time : 5.4 minutes                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> [1] "Index directory already exists remove it if you want to recreate it"
#> 
#>         ==========     _____ _    _ ____  _____  ______          _____  
#>         =====         / ____| |  | |  _ \|  __ \|  ____|   /\   |  __ \ 
#>           =====      | (___ | |  | | |_) | |__) | |__     /  \  | |  | |
#>             ====      \___ \| |  | |  _ <|  _  /|  __|   / /\ \ | |  | |
#>               ====    ____) | |__| | |_) | | \ \| |____ / ____ \| |__| |
#>         ==========   |_____/ \____/|____/|_|  \_\______/_/    \_\_____/
#>        Rsubread 2.20.0
#> 
#> //================================= setting ==================================\\
#> ||                                                                            ||
#> || Function      : Read alignment (DNA-Seq)                                   ||
#> || Input file    : prepro.PPP-BG.10M_CA_valid.fastq.gz                        ||
#> || Output file   : prepro.PPP-BG.10M_CA_valid.fastq.gz.bam (BAM), Sorted      ||
#> || Index name    : index                                                      ||
#> ||                                                                            ||
#> ||                    ------------------------------------                    ||
#> ||                                                                            ||
#> ||                               Threads : 1                                  ||
#> ||                          Phred offset : 33                                 ||
#> ||                             Min votes : 3 / 10                             ||
#> ||                        Max mismatches : 1                                  ||
#> ||                      Max indel length : 0                                  ||
#> ||            Report multi-mapping reads : no                                 ||
#> || Max alignments per multi-mapping read : 1                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> //================ Running (01-Nov-2025 18:19:40, pid=392792) ================\\
#> ||                                                                            ||
#> || Check the input reads.                                                     ||
#> || The input file contains base space reads.                                  ||
#> || Initialise the memory objects.                                             ||
#> || Estimate the mean read length.                                             ||
#> || WARNING  - The specified Phred score offset (33) seems incorrect.          ||
#> ||            ASCII values of the quality scores of read bases included in    ||
#> ||            the first 3000 reads were found to be within the range of       ||
#> ||            [63,63].                                                        ||
#> ||                                                                            ||
#> || Create the output BAM file.                                                ||
#> || Check the index.                                                           ||
#> || Init the voting space.                                                     ||
#> || Global environment is initialised.                                         ||
#> || Load the 1-th index block...                                               ||
#> || The index block has been loaded.                                           ||
#> || Start read mapping in chunk.                                               ||
#> ||    0% completed, 1.2 mins elapsed, rate=102.8k reads per second            ||
#> ||    6% completed, 1.4 mins elapsed, rate=111.8k reads per second            ||
#> ||   13% completed, 1.6 mins elapsed, rate=111.9k reads per second            ||
#> ||   20% completed, 1.8 mins elapsed, rate=112.2k reads per second            ||
#> ||   24% completed, 1.9 mins elapsed, rate=42.4k reads per second             ||
#> ||   25% completed, 2.0 mins elapsed, rate=43.0k reads per second             ||
#> ||   26% completed, 2.1 mins elapsed, rate=43.5k reads per second             ||
#> ||   27% completed, 2.1 mins elapsed, rate=44.0k reads per second             ||
#> ||   29% completed, 2.2 mins elapsed, rate=44.5k reads per second             ||
#> ||   30% completed, 2.2 mins elapsed, rate=45.0k reads per second             ||
#> ||   31% completed, 2.3 mins elapsed, rate=45.4k reads per second             ||
#> ||   32% completed, 2.4 mins elapsed, rate=45.9k reads per second             ||
#> ||   33% completed, 2.4 mins elapsed, rate=46.3k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   34% completed, 2.6 mins elapsed, rate=81.5k reads per second             ||
#> ||   41% completed, 2.8 mins elapsed, rate=85.6k reads per second             ||
#> ||   48% completed, 3.0 mins elapsed, rate=88.9k reads per second             ||
#> ||   54% completed, 3.2 mins elapsed, rate=91.5k reads per second             ||
#> ||   59% completed, 3.3 mins elapsed, rate=59.4k reads per second             ||
#> ||   60% completed, 3.4 mins elapsed, rate=59.5k reads per second             ||
#> ||   61% completed, 3.5 mins elapsed, rate=59.5k reads per second             ||
#> ||   62% completed, 3.5 mins elapsed, rate=59.6k reads per second             ||
#> ||   64% completed, 3.6 mins elapsed, rate=59.6k reads per second             ||
#> ||   65% completed, 3.6 mins elapsed, rate=59.6k reads per second             ||
#> ||   66% completed, 3.7 mins elapsed, rate=59.7k reads per second             ||
#> ||   67% completed, 3.8 mins elapsed, rate=59.7k reads per second             ||
#> ||   68% completed, 3.8 mins elapsed, rate=59.8k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   69% completed, 4.0 mins elapsed, rate=83.2k reads per second             ||
#> ||   76% completed, 4.2 mins elapsed, rate=85.3k reads per second             ||
#> ||   83% completed, 4.4 mins elapsed, rate=87.1k reads per second             ||
#> ||   89% completed, 4.5 mins elapsed, rate=88.7k reads per second             ||
#> ||   90% completed, 4.6 mins elapsed, rate=65.9k reads per second             ||
#> ||   91% completed, 4.7 mins elapsed, rate=65.8k reads per second             ||
#> ||   92% completed, 4.7 mins elapsed, rate=65.8k reads per second             ||
#> ||   93% completed, 4.8 mins elapsed, rate=65.8k reads per second             ||
#> ||   94% completed, 4.8 mins elapsed, rate=65.7k reads per second             ||
#> ||   96% completed, 4.9 mins elapsed, rate=65.7k reads per second             ||
#> ||   97% completed, 4.9 mins elapsed, rate=65.6k reads per second             ||
#> ||   98% completed, 5.0 mins elapsed, rate=65.6k reads per second             ||
#> ||   99% completed, 5.0 mins elapsed, rate=65.6k reads per second             ||
#> ||                                                                            ||
#> ||                           Completed successfully.                          ||
#> ||                                                                            ||
#> \\====================================    ====================================//
#> 
#> //================================   Summary =================================\\
#> ||                                                                            ||
#> ||                 Total reads : 20,000,000                                   ||
#> ||                      Mapped : 3,747,824 (18.7%)                            ||
#> ||             Uniquely mapped : 3,747,824                                    ||
#> ||               Multi-mapping : 0                                            ||
#> ||                                                                            ||
#> ||                    Unmapped : 16,252,176                                   ||
#> ||                                                                            ||
#> ||                      Indels : 0                                            ||
#> ||                                                                            ||
#> ||                     WARNING : Phred offset (33) incorrect?                 ||
#> ||                                                                            ||
#> ||                Running time : 5.6 minutes                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> [1] "Index directory already exists remove it if you want to recreate it"
#> 
#>         ==========     _____ _    _ ____  _____  ______          _____  
#>         =====         / ____| |  | |  _ \|  __ \|  ____|   /\   |  __ \ 
#>           =====      | (___ | |  | | |_) | |__) | |__     /  \  | |  | |
#>             ====      \___ \| |  | |  _ <|  _  /|  __|   / /\ \ | |  | |
#>               ====    ____) | |__| | |_) | | \ \| |____ / ____ \| |__| |
#>         ==========   |_____/ \____/|____/|_|  \_\______/_/    \_\_____/
#>        Rsubread 2.20.0
#> 
#> //================================= setting ==================================\\
#> ||                                                                            ||
#> || Function      : Read alignment (DNA-Seq)                                   ||
#> || Input file    : prepro.PPP-BG.10M_CC_valid.fastq.gz                        ||
#> || Output file   : prepro.PPP-BG.10M_CC_valid.fastq.gz.bam (BAM), Sorted      ||
#> || Index name    : index                                                      ||
#> ||                                                                            ||
#> ||                    ------------------------------------                    ||
#> ||                                                                            ||
#> ||                               Threads : 1                                  ||
#> ||                          Phred offset : 33                                 ||
#> ||                             Min votes : 3 / 10                             ||
#> ||                        Max mismatches : 1                                  ||
#> ||                      Max indel length : 0                                  ||
#> ||            Report multi-mapping reads : no                                 ||
#> || Max alignments per multi-mapping read : 1                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> //================ Running (01-Nov-2025 18:25:17, pid=392792) ================\\
#> ||                                                                            ||
#> || Check the input reads.                                                     ||
#> || The input file contains base space reads.                                  ||
#> || Initialise the memory objects.                                             ||
#> || Estimate the mean read length.                                             ||
#> || WARNING  - The specified Phred score offset (33) seems incorrect.          ||
#> ||            ASCII values of the quality scores of read bases included in    ||
#> ||            the first 3000 reads were found to be within the range of       ||
#> ||            [63,63].                                                        ||
#> ||                                                                            ||
#> || Create the output BAM file.                                                ||
#> || Check the index.                                                           ||
#> || Init the voting space.                                                     ||
#> || Global environment is initialised.                                         ||
#> || Load the 1-th index block...                                               ||
#> || The index block has been loaded.                                           ||
#> || Start read mapping in chunk.                                               ||
#> ||    0% completed, 1.2 mins elapsed, rate=103.1k reads per second            ||
#> ||    6% completed, 1.4 mins elapsed, rate=113.0k reads per second            ||
#> ||   13% completed, 1.6 mins elapsed, rate=112.8k reads per second            ||
#> ||   19% completed, 1.7 mins elapsed, rate=112.5k reads per second            ||
#> ||   24% completed, 1.9 mins elapsed, rate=42.5k reads per second             ||
#> ||   25% completed, 2.0 mins elapsed, rate=43.1k reads per second             ||
#> ||   26% completed, 2.0 mins elapsed, rate=43.6k reads per second             ||
#> ||   27% completed, 2.1 mins elapsed, rate=44.1k reads per second             ||
#> ||   29% completed, 2.2 mins elapsed, rate=44.6k reads per second             ||
#> ||   30% completed, 2.2 mins elapsed, rate=45.1k reads per second             ||
#> ||   31% completed, 2.3 mins elapsed, rate=45.5k reads per second             ||
#> ||   32% completed, 2.4 mins elapsed, rate=45.9k reads per second             ||
#> ||   33% completed, 2.4 mins elapsed, rate=46.3k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   35% completed, 2.6 mins elapsed, rate=82.2k reads per second             ||
#> ||   41% completed, 2.8 mins elapsed, rate=86.0k reads per second             ||
#> ||   48% completed, 3.0 mins elapsed, rate=89.2k reads per second             ||
#> ||   55% completed, 3.2 mins elapsed, rate=91.8k reads per second             ||
#> ||   59% completed, 3.3 mins elapsed, rate=59.7k reads per second             ||
#> ||   60% completed, 3.4 mins elapsed, rate=59.8k reads per second             ||
#> ||   61% completed, 3.4 mins elapsed, rate=59.8k reads per second             ||
#> ||   62% completed, 3.5 mins elapsed, rate=59.9k reads per second             ||
#> ||   64% completed, 3.6 mins elapsed, rate=59.9k reads per second             ||
#> ||   65% completed, 3.6 mins elapsed, rate=60.0k reads per second             ||
#> ||   66% completed, 3.7 mins elapsed, rate=60.0k reads per second             ||
#> ||   67% completed, 3.8 mins elapsed, rate=60.0k reads per second             ||
#> ||   68% completed, 3.8 mins elapsed, rate=60.1k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   69% completed, 4.0 mins elapsed, rate=83.4k reads per second             ||
#> ||   76% completed, 4.1 mins elapsed, rate=85.5k reads per second             ||
#> ||   83% completed, 4.3 mins elapsed, rate=87.4k reads per second             ||
#> ||   89% completed, 4.5 mins elapsed, rate=89.1k reads per second             ||
#> ||   90% completed, 4.6 mins elapsed, rate=66.1k reads per second             ||
#> ||   91% completed, 4.6 mins elapsed, rate=66.1k reads per second             ||
#> ||   92% completed, 4.7 mins elapsed, rate=66.0k reads per second             ||
#> ||   93% completed, 4.7 mins elapsed, rate=66.0k reads per second             ||
#> ||   94% completed, 4.8 mins elapsed, rate=65.9k reads per second             ||
#> ||   95% completed, 4.9 mins elapsed, rate=65.9k reads per second             ||
#> ||   96% completed, 4.9 mins elapsed, rate=65.8k reads per second             ||
#> ||   97% completed, 5.0 mins elapsed, rate=65.8k reads per second             ||
#> ||   98% completed, 5.0 mins elapsed, rate=65.7k reads per second             ||
#> ||                                                                            ||
#> ||                           Completed successfully.                          ||
#> ||                                                                            ||
#> \\====================================    ====================================//
#> 
#> //================================   Summary =================================\\
#> ||                                                                            ||
#> ||                 Total reads : 20,000,000                                   ||
#> ||                      Mapped : 3,751,602 (18.8%)                            ||
#> ||             Uniquely mapped : 3,751,602                                    ||
#> ||               Multi-mapping : 0                                            ||
#> ||                                                                            ||
#> ||                    Unmapped : 16,248,398                                   ||
#> ||                                                                            ||
#> ||                      Indels : 0                                            ||
#> ||                                                                            ||
#> ||                     WARNING : Phred offset (33) incorrect?                 ||
#> ||                                                                            ||
#> ||                Running time : 5.6 minutes                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> [1] "Index directory already exists remove it if you want to recreate it"
#> 
#>         ==========     _____ _    _ ____  _____  ______          _____  
#>         =====         / ____| |  | |  _ \|  __ \|  ____|   /\   |  __ \ 
#>           =====      | (___ | |  | | |_) | |__) | |__     /  \  | |  | |
#>             ====      \___ \| |  | |  _ <|  _  /|  __|   / /\ \ | |  | |
#>               ====    ____) | |__| | |_) | | \ \| |____ / ____ \| |__| |
#>         ==========   |_____/ \____/|____/|_|  \_\______/_/    \_\_____/
#>        Rsubread 2.20.0
#> 
#> //================================= setting ==================================\\
#> ||                                                                            ||
#> || Function      : Read alignment (DNA-Seq)                                   ||
#> || Input file    : prepro.PPP-BG.10M_CT_valid.fastq.gz                        ||
#> || Output file   : prepro.PPP-BG.10M_CT_valid.fastq.gz.bam (BAM), Sorted      ||
#> || Index name    : index                                                      ||
#> ||                                                                            ||
#> ||                    ------------------------------------                    ||
#> ||                                                                            ||
#> ||                               Threads : 1                                  ||
#> ||                          Phred offset : 33                                 ||
#> ||                             Min votes : 3 / 10                             ||
#> ||                        Max mismatches : 1                                  ||
#> ||                      Max indel length : 0                                  ||
#> ||            Report multi-mapping reads : no                                 ||
#> || Max alignments per multi-mapping read : 1                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> //================ Running (01-Nov-2025 18:30:52, pid=392792) ================\\
#> ||                                                                            ||
#> || Check the input reads.                                                     ||
#> || The input file contains base space reads.                                  ||
#> || Initialise the memory objects.                                             ||
#> || Estimate the mean read length.                                             ||
#> || WARNING  - The specified Phred score offset (33) seems incorrect.          ||
#> ||            ASCII values of the quality scores of read bases included in    ||
#> ||            the first 3000 reads were found to be within the range of       ||
#> ||            [63,63].                                                        ||
#> ||                                                                            ||
#> || Create the output BAM file.                                                ||
#> || Check the index.                                                           ||
#> || Init the voting space.                                                     ||
#> || Global environment is initialised.                                         ||
#> || Load the 1-th index block...                                               ||
#> || The index block has been loaded.                                           ||
#> || Start read mapping in chunk.                                               ||
#> ||    0% completed, 1.2 mins elapsed, rate=101.1k reads per second            ||
#> ||    6% completed, 1.4 mins elapsed, rate=112.6k reads per second            ||
#> ||   13% completed, 1.6 mins elapsed, rate=112.5k reads per second            ||
#> ||   20% completed, 1.8 mins elapsed, rate=112.5k reads per second            ||
#> ||   24% completed, 1.9 mins elapsed, rate=42.5k reads per second             ||
#> ||   25% completed, 2.0 mins elapsed, rate=43.1k reads per second             ||
#> ||   26% completed, 2.0 mins elapsed, rate=43.6k reads per second             ||
#> ||   27% completed, 2.1 mins elapsed, rate=44.1k reads per second             ||
#> ||   28% completed, 2.2 mins elapsed, rate=44.6k reads per second             ||
#> ||   30% completed, 2.2 mins elapsed, rate=45.1k reads per second             ||
#> ||   31% completed, 2.3 mins elapsed, rate=45.5k reads per second             ||
#> ||   32% completed, 2.4 mins elapsed, rate=45.8k reads per second             ||
#> ||   33% completed, 2.4 mins elapsed, rate=46.2k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   34% completed, 2.6 mins elapsed, rate=82.0k reads per second             ||
#> ||   41% completed, 2.8 mins elapsed, rate=86.0k reads per second             ||
#> ||   48% completed, 3.0 mins elapsed, rate=89.3k reads per second             ||
#> ||   54% completed, 3.2 mins elapsed, rate=91.9k reads per second             ||
#> ||   59% completed, 3.3 mins elapsed, rate=59.6k reads per second             ||
#> ||   60% completed, 3.4 mins elapsed, rate=59.7k reads per second             ||
#> ||   61% completed, 3.4 mins elapsed, rate=59.7k reads per second             ||
#> ||   62% completed, 3.5 mins elapsed, rate=59.8k reads per second             ||
#> ||   63% completed, 3.6 mins elapsed, rate=59.8k reads per second             ||
#> ||   65% completed, 3.6 mins elapsed, rate=59.9k reads per second             ||
#> ||   66% completed, 3.7 mins elapsed, rate=59.9k reads per second             ||
#> ||   67% completed, 3.8 mins elapsed, rate=59.9k reads per second             ||
#> ||   68% completed, 3.8 mins elapsed, rate=60.0k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   69% completed, 4.0 mins elapsed, rate=83.6k reads per second             ||
#> ||   76% completed, 4.1 mins elapsed, rate=85.7k reads per second             ||
#> ||   83% completed, 4.3 mins elapsed, rate=87.5k reads per second             ||
#> ||   89% completed, 4.5 mins elapsed, rate=89.2k reads per second             ||
#> ||   90% completed, 4.6 mins elapsed, rate=66.1k reads per second             ||
#> ||   91% completed, 4.6 mins elapsed, rate=66.1k reads per second             ||
#> ||   92% completed, 4.7 mins elapsed, rate=66.0k reads per second             ||
#> ||   93% completed, 4.7 mins elapsed, rate=66.0k reads per second             ||
#> ||   94% completed, 4.8 mins elapsed, rate=65.9k reads per second             ||
#> ||   95% completed, 4.9 mins elapsed, rate=65.9k reads per second             ||
#> ||   96% completed, 4.9 mins elapsed, rate=65.9k reads per second             ||
#> ||   97% completed, 5.0 mins elapsed, rate=65.8k reads per second             ||
#> ||   98% completed, 5.0 mins elapsed, rate=65.8k reads per second             ||
#> ||                                                                            ||
#> ||                           Completed successfully.                          ||
#> ||                                                                            ||
#> \\====================================    ====================================//
#> 
#> //================================   Summary =================================\\
#> ||                                                                            ||
#> ||                 Total reads : 20,000,000                                   ||
#> ||                      Mapped : 3,750,703 (18.8%)                            ||
#> ||             Uniquely mapped : 3,750,703                                    ||
#> ||               Multi-mapping : 0                                            ||
#> ||                                                                            ||
#> ||                    Unmapped : 16,249,297                                   ||
#> ||                                                                            ||
#> ||                      Indels : 0                                            ||
#> ||                                                                            ||
#> ||                     WARNING : Phred offset (33) incorrect?                 ||
#> ||                                                                            ||
#> ||                Running time : 5.6 minutes                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> [1] "Index directory already exists remove it if you want to recreate it"
#> 
#>         ==========     _____ _    _ ____  _____  ______          _____  
#>         =====         / ____| |  | |  _ \|  __ \|  ____|   /\   |  __ \ 
#>           =====      | (___ | |  | | |_) | |__) | |__     /  \  | |  | |
#>             ====      \___ \| |  | |  _ <|  _  /|  __|   / /\ \ | |  | |
#>               ====    ____) | |__| | |_) | | \ \| |____ / ____ \| |__| |
#>         ==========   |_____/ \____/|____/|_|  \_\______/_/    \_\_____/
#>        Rsubread 2.20.0
#> 
#> //================================= setting ==================================\\
#> ||                                                                            ||
#> || Function      : Read alignment (DNA-Seq)                                   ||
#> || Input file    : prepro.PPP-BG.15M_AA_valid.fastq.gz                        ||
#> || Output file   : prepro.PPP-BG.15M_AA_valid.fastq.gz.bam (BAM), Sorted      ||
#> || Index name    : index                                                      ||
#> ||                                                                            ||
#> ||                    ------------------------------------                    ||
#> ||                                                                            ||
#> ||                               Threads : 1                                  ||
#> ||                          Phred offset : 33                                 ||
#> ||                             Min votes : 3 / 10                             ||
#> ||                        Max mismatches : 1                                  ||
#> ||                      Max indel length : 0                                  ||
#> ||            Report multi-mapping reads : no                                 ||
#> || Max alignments per multi-mapping read : 1                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> //================ Running (01-Nov-2025 18:36:31, pid=392792) ================\\
#> ||                                                                            ||
#> || Check the input reads.                                                     ||
#> || The input file contains base space reads.                                  ||
#> || Initialise the memory objects.                                             ||
#> || Estimate the mean read length.                                             ||
#> || WARNING  - The specified Phred score offset (33) seems incorrect.          ||
#> ||            ASCII values of the quality scores of read bases included in    ||
#> ||            the first 3000 reads were found to be within the range of       ||
#> ||            [63,63].                                                        ||
#> ||                                                                            ||
#> || Create the output BAM file.                                                ||
#> || Check the index.                                                           ||
#> || Init the voting space.                                                     ||
#> || Global environment is initialised.                                         ||
#> || Load the 1-th index block...                                               ||
#> || The index block has been loaded.                                           ||
#> || Start read mapping in chunk.                                               ||
#> ||    0% completed, 1.2 mins elapsed, rate=96.8k reads per second             ||
#> ||    6% completed, 1.4 mins elapsed, rate=106.3k reads per second            ||
#> ||   12% completed, 1.8 mins elapsed, rate=106.7k reads per second            ||
#> ||   16% completed, 1.9 mins elapsed, rate=41.9k reads per second             ||
#> ||   17% completed, 2.0 mins elapsed, rate=42.7k reads per second             ||
#> ||   17% completed, 2.1 mins elapsed, rate=43.5k reads per second             ||
#> ||   18% completed, 2.1 mins elapsed, rate=44.2k reads per second             ||
#> ||   19% completed, 2.2 mins elapsed, rate=44.9k reads per second             ||
#> ||   20% completed, 2.2 mins elapsed, rate=45.6k reads per second             ||
#> ||   20% completed, 2.3 mins elapsed, rate=46.2k reads per second             ||
#> ||   21% completed, 2.3 mins elapsed, rate=46.8k reads per second             ||
#> ||   22% completed, 2.4 mins elapsed, rate=47.4k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   23% completed, 2.6 mins elapsed, rate=83.4k reads per second             ||
#> ||   29% completed, 2.9 mins elapsed, rate=88.3k reads per second             ||
#> ||   36% completed, 3.2 mins elapsed, rate=91.6k reads per second             ||
#> ||   39% completed, 3.3 mins elapsed, rate=59.7k reads per second             ||
#> ||   40% completed, 3.4 mins elapsed, rate=59.9k reads per second             ||
#> ||   41% completed, 3.4 mins elapsed, rate=60.1k reads per second             ||
#> ||   41% completed, 3.5 mins elapsed, rate=60.4k reads per second             ||
#> ||   42% completed, 3.5 mins elapsed, rate=60.5k reads per second             ||
#> ||   43% completed, 3.6 mins elapsed, rate=60.8k reads per second             ||
#> ||   44% completed, 3.6 mins elapsed, rate=60.9k reads per second             ||
#> ||   44% completed, 3.7 mins elapsed, rate=61.1k reads per second             ||
#> ||   45% completed, 3.7 mins elapsed, rate=61.3k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   46% completed, 3.9 mins elapsed, rate=84.9k reads per second             ||
#> ||   53% completed, 4.2 mins elapsed, rate=87.4k reads per second             ||
#> ||   59% completed, 4.5 mins elapsed, rate=89.4k reads per second             ||
#> ||   62% completed, 4.7 mins elapsed, rate=67.4k reads per second             ||
#> ||   63% completed, 4.7 mins elapsed, rate=67.5k reads per second             ||
#> ||   64% completed, 4.8 mins elapsed, rate=67.5k reads per second             ||
#> ||   65% completed, 4.8 mins elapsed, rate=67.6k reads per second             ||
#> ||   65% completed, 4.9 mins elapsed, rate=67.7k reads per second             ||
#> ||   66% completed, 4.9 mins elapsed, rate=67.8k reads per second             ||
#> ||   67% completed, 5.0 mins elapsed, rate=67.8k reads per second             ||
#> ||   68% completed, 5.0 mins elapsed, rate=67.9k reads per second             ||
#> ||   69% completed, 5.1 mins elapsed, rate=68.0k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   69% completed, 5.3 mins elapsed, rate=85.1k reads per second             ||
#> ||   76% completed, 5.6 mins elapsed, rate=86.9k reads per second             ||
#> ||   83% completed, 5.9 mins elapsed, rate=88.4k reads per second             ||
#> ||   86% completed, 6.0 mins elapsed, rate=71.5k reads per second             ||
#> ||   86% completed, 6.1 mins elapsed, rate=71.6k reads per second             ||
#> ||   87% completed, 6.1 mins elapsed, rate=71.6k reads per second             ||
#> ||   88% completed, 6.2 mins elapsed, rate=71.6k reads per second             ||
#> ||   89% completed, 6.2 mins elapsed, rate=71.6k reads per second             ||
#> ||   89% completed, 6.3 mins elapsed, rate=71.6k reads per second             ||
#> ||   90% completed, 6.3 mins elapsed, rate=71.6k reads per second             ||
#> ||   91% completed, 6.4 mins elapsed, rate=71.7k reads per second             ||
#> ||   92% completed, 6.4 mins elapsed, rate=71.7k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   93% completed, 6.6 mins elapsed, rate=85.4k reads per second             ||
#> ||   97% completed, 6.8 mins elapsed, rate=71.6k reads per second             ||
#> ||   98% completed, 6.9 mins elapsed, rate=71.6k reads per second             ||
#> ||   98% completed, 6.9 mins elapsed, rate=71.6k reads per second             ||
#> ||   98% completed, 6.9 mins elapsed, rate=71.6k reads per second             ||
#> ||   98% completed, 6.9 mins elapsed, rate=71.6k reads per second             ||
#> ||   99% completed, 6.9 mins elapsed, rate=71.6k reads per second             ||
#> ||   99% completed, 6.9 mins elapsed, rate=71.6k reads per second             ||
#> ||   99% completed, 7.0 mins elapsed, rate=71.6k reads per second             ||
#> ||   99% completed, 7.0 mins elapsed, rate=71.6k reads per second             ||
#> ||                                                                            ||
#> ||                           Completed successfully.                          ||
#> ||                                                                            ||
#> \\====================================    ====================================//
#> 
#> //================================   Summary =================================\\
#> ||                                                                            ||
#> ||                 Total reads : 30,000,000                                   ||
#> ||                      Mapped : 21,355,573 (71.2%)                           ||
#> ||             Uniquely mapped : 21,355,573                                   ||
#> ||               Multi-mapping : 0                                            ||
#> ||                                                                            ||
#> ||                    Unmapped : 8,644,427                                    ||
#> ||                                                                            ||
#> ||                      Indels : 0                                            ||
#> ||                                                                            ||
#> ||                     WARNING : Phred offset (33) incorrect?                 ||
#> ||                                                                            ||
#> ||                Running time : 7.7 minutes                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> [1] "Index directory already exists remove it if you want to recreate it"
#> 
#>         ==========     _____ _    _ ____  _____  ______          _____  
#>         =====         / ____| |  | |  _ \|  __ \|  ____|   /\   |  __ \ 
#>           =====      | (___ | |  | | |_) | |__) | |__     /  \  | |  | |
#>             ====      \___ \| |  | |  _ <|  _  /|  __|   / /\ \ | |  | |
#>               ====    ____) | |__| | |_) | | \ \| |____ / ____ \| |__| |
#>         ==========   |_____/ \____/|____/|_|  \_\______/_/    \_\_____/
#>        Rsubread 2.20.0
#> 
#> //================================= setting ==================================\\
#> ||                                                                            ||
#> || Function      : Read alignment (DNA-Seq)                                   ||
#> || Input file    : prepro.PPP-BG.15M_AC_valid.fastq.gz                        ||
#> || Output file   : prepro.PPP-BG.15M_AC_valid.fastq.gz.bam (BAM), Sorted      ||
#> || Index name    : index                                                      ||
#> ||                                                                            ||
#> ||                    ------------------------------------                    ||
#> ||                                                                            ||
#> ||                               Threads : 1                                  ||
#> ||                          Phred offset : 33                                 ||
#> ||                             Min votes : 3 / 10                             ||
#> ||                        Max mismatches : 1                                  ||
#> ||                      Max indel length : 0                                  ||
#> ||            Report multi-mapping reads : no                                 ||
#> || Max alignments per multi-mapping read : 1                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> //================ Running (01-Nov-2025 18:44:13, pid=392792) ================\\
#> ||                                                                            ||
#> || Check the input reads.                                                     ||
#> || The input file contains base space reads.                                  ||
#> || Initialise the memory objects.                                             ||
#> || Estimate the mean read length.                                             ||
#> || WARNING  - The specified Phred score offset (33) seems incorrect.          ||
#> ||            ASCII values of the quality scores of read bases included in    ||
#> ||            the first 3000 reads were found to be within the range of       ||
#> ||            [63,63].                                                        ||
#> ||                                                                            ||
#> || Create the output BAM file.                                                ||
#> || Check the index.                                                           ||
#> || Init the voting space.                                                     ||
#> || Global environment is initialised.                                         ||
#> || Load the 1-th index block...                                               ||
#> || The index block has been loaded.                                           ||
#> || Start read mapping in chunk.                                               ||
#> ||    0% completed, 1.2 mins elapsed, rate=101.8k reads per second            ||
#> ||    6% completed, 1.5 mins elapsed, rate=106.9k reads per second            ||
#> ||   12% completed, 1.8 mins elapsed, rate=107.0k reads per second            ||
#> ||   16% completed, 1.9 mins elapsed, rate=42.0k reads per second             ||
#> ||   17% completed, 2.0 mins elapsed, rate=42.7k reads per second             ||
#> ||   17% completed, 2.1 mins elapsed, rate=43.5k reads per second             ||
#> ||   18% completed, 2.1 mins elapsed, rate=44.2k reads per second             ||
#> ||   19% completed, 2.2 mins elapsed, rate=44.9k reads per second             ||
#> ||   20% completed, 2.2 mins elapsed, rate=45.6k reads per second             ||
#> ||   20% completed, 2.3 mins elapsed, rate=46.2k reads per second             ||
#> ||   21% completed, 2.3 mins elapsed, rate=46.8k reads per second             ||
#> ||   22% completed, 2.4 mins elapsed, rate=47.4k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   23% completed, 2.6 mins elapsed, rate=83.6k reads per second             ||
#> ||   29% completed, 2.9 mins elapsed, rate=88.4k reads per second             ||
#> ||   36% completed, 3.2 mins elapsed, rate=91.9k reads per second             ||
#> ||   39% completed, 3.3 mins elapsed, rate=59.8k reads per second             ||
#> ||   40% completed, 3.4 mins elapsed, rate=60.0k reads per second             ||
#> ||   41% completed, 3.4 mins elapsed, rate=60.2k reads per second             ||
#> ||   41% completed, 3.5 mins elapsed, rate=60.4k reads per second             ||
#> ||   42% completed, 3.5 mins elapsed, rate=60.6k reads per second             ||
#> ||   43% completed, 3.6 mins elapsed, rate=60.8k reads per second             ||
#> ||   44% completed, 3.6 mins elapsed, rate=61.0k reads per second             ||
#> ||   44% completed, 3.7 mins elapsed, rate=61.2k reads per second             ||
#> ||   45% completed, 3.7 mins elapsed, rate=61.4k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   46% completed, 3.9 mins elapsed, rate=85.0k reads per second             ||
#> ||   53% completed, 4.2 mins elapsed, rate=87.5k reads per second             ||
#> ||   59% completed, 4.5 mins elapsed, rate=89.6k reads per second             ||
#> ||   62% completed, 4.7 mins elapsed, rate=67.4k reads per second             ||
#> ||   63% completed, 4.7 mins elapsed, rate=67.5k reads per second             ||
#> ||   64% completed, 4.8 mins elapsed, rate=67.6k reads per second             ||
#> ||   65% completed, 4.8 mins elapsed, rate=67.6k reads per second             ||
#> ||   65% completed, 4.9 mins elapsed, rate=67.7k reads per second             ||
#> ||   66% completed, 4.9 mins elapsed, rate=67.8k reads per second             ||
#> ||   67% completed, 5.0 mins elapsed, rate=67.8k reads per second             ||
#> ||   68% completed, 5.0 mins elapsed, rate=67.9k reads per second             ||
#> ||   69% completed, 5.1 mins elapsed, rate=68.0k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   69% completed, 5.3 mins elapsed, rate=85.4k reads per second             ||
#> ||   76% completed, 5.6 mins elapsed, rate=87.1k reads per second             ||
#> ||   83% completed, 5.9 mins elapsed, rate=88.7k reads per second             ||
#> ||   86% completed, 6.0 mins elapsed, rate=71.8k reads per second             ||
#> ||   86% completed, 6.1 mins elapsed, rate=71.8k reads per second             ||
#> ||   87% completed, 6.1 mins elapsed, rate=71.8k reads per second             ||
#> ||   88% completed, 6.2 mins elapsed, rate=71.9k reads per second             ||
#> ||   89% completed, 6.2 mins elapsed, rate=71.9k reads per second             ||
#> ||   90% completed, 6.3 mins elapsed, rate=71.9k reads per second             ||
#> ||   90% completed, 6.3 mins elapsed, rate=72.0k reads per second             ||
#> ||   91% completed, 6.4 mins elapsed, rate=72.0k reads per second             ||
#> ||   92% completed, 6.4 mins elapsed, rate=72.0k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   93% completed, 6.6 mins elapsed, rate=85.9k reads per second             ||
#> ||   97% completed, 6.8 mins elapsed, rate=72.0k reads per second             ||
#> ||   98% completed, 6.8 mins elapsed, rate=72.0k reads per second             ||
#> ||   98% completed, 6.8 mins elapsed, rate=72.0k reads per second             ||
#> ||   98% completed, 6.9 mins elapsed, rate=72.0k reads per second             ||
#> ||   98% completed, 6.9 mins elapsed, rate=72.0k reads per second             ||
#> ||   99% completed, 6.9 mins elapsed, rate=72.0k reads per second             ||
#> ||   99% completed, 6.9 mins elapsed, rate=72.0k reads per second             ||
#> ||   99% completed, 6.9 mins elapsed, rate=72.0k reads per second             ||
#> ||   99% completed, 6.9 mins elapsed, rate=72.0k reads per second             ||
#> ||                                                                            ||
#> ||                           Completed successfully.                          ||
#> ||                                                                            ||
#> \\====================================    ====================================//
#> 
#> //================================   Summary =================================\\
#> ||                                                                            ||
#> ||                 Total reads : 30,000,000                                   ||
#> ||                      Mapped : 21,351,406 (71.2%)                           ||
#> ||             Uniquely mapped : 21,351,406                                   ||
#> ||               Multi-mapping : 0                                            ||
#> ||                                                                            ||
#> ||                    Unmapped : 8,648,594                                    ||
#> ||                                                                            ||
#> ||                      Indels : 0                                            ||
#> ||                                                                            ||
#> ||                     WARNING : Phred offset (33) incorrect?                 ||
#> ||                                                                            ||
#> ||                Running time : 7.6 minutes                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> [1] "Index directory already exists remove it if you want to recreate it"
#> 
#>         ==========     _____ _    _ ____  _____  ______          _____  
#>         =====         / ____| |  | |  _ \|  __ \|  ____|   /\   |  __ \ 
#>           =====      | (___ | |  | | |_) | |__) | |__     /  \  | |  | |
#>             ====      \___ \| |  | |  _ <|  _  /|  __|   / /\ \ | |  | |
#>               ====    ____) | |__| | |_) | | \ \| |____ / ____ \| |__| |
#>         ==========   |_____/ \____/|____/|_|  \_\______/_/    \_\_____/
#>        Rsubread 2.20.0
#> 
#> //================================= setting ==================================\\
#> ||                                                                            ||
#> || Function      : Read alignment (DNA-Seq)                                   ||
#> || Input file    : prepro.PPP-BG.15M_AT_valid.fastq.gz                        ||
#> || Output file   : prepro.PPP-BG.15M_AT_valid.fastq.gz.bam (BAM), Sorted      ||
#> || Index name    : index                                                      ||
#> ||                                                                            ||
#> ||                    ------------------------------------                    ||
#> ||                                                                            ||
#> ||                               Threads : 1                                  ||
#> ||                          Phred offset : 33                                 ||
#> ||                             Min votes : 3 / 10                             ||
#> ||                        Max mismatches : 1                                  ||
#> ||                      Max indel length : 0                                  ||
#> ||            Report multi-mapping reads : no                                 ||
#> || Max alignments per multi-mapping read : 1                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> //================ Running (01-Nov-2025 18:51:50, pid=392792) ================\\
#> ||                                                                            ||
#> || Check the input reads.                                                     ||
#> || The input file contains base space reads.                                  ||
#> || Initialise the memory objects.                                             ||
#> || Estimate the mean read length.                                             ||
#> || WARNING  - The specified Phred score offset (33) seems incorrect.          ||
#> ||            ASCII values of the quality scores of read bases included in    ||
#> ||            the first 3000 reads were found to be within the range of       ||
#> ||            [63,63].                                                        ||
#> ||                                                                            ||
#> || Create the output BAM file.                                                ||
#> || Check the index.                                                           ||
#> || Init the voting space.                                                     ||
#> || Global environment is initialised.                                         ||
#> || Load the 1-th index block...                                               ||
#> || The index block has been loaded.                                           ||
#> || Start read mapping in chunk.                                               ||
#> ||    0% completed, 1.2 mins elapsed, rate=75.0k reads per second             ||
#> ||    6% completed, 1.4 mins elapsed, rate=108.6k reads per second            ||
#> ||   12% completed, 1.7 mins elapsed, rate=108.9k reads per second            ||
#> ||   16% completed, 1.9 mins elapsed, rate=42.5k reads per second             ||
#> ||   17% completed, 2.0 mins elapsed, rate=43.3k reads per second             ||
#> ||   17% completed, 2.0 mins elapsed, rate=44.1k reads per second             ||
#> ||   18% completed, 2.1 mins elapsed, rate=44.8k reads per second             ||
#> ||   19% completed, 2.1 mins elapsed, rate=45.5k reads per second             ||
#> ||   20% completed, 2.2 mins elapsed, rate=46.2k reads per second             ||
#> ||   20% completed, 2.2 mins elapsed, rate=46.8k reads per second             ||
#> ||   21% completed, 2.3 mins elapsed, rate=47.4k reads per second             ||
#> ||   22% completed, 2.3 mins elapsed, rate=48.0k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   23% completed, 2.5 mins elapsed, rate=85.3k reads per second             ||
#> ||   29% completed, 2.8 mins elapsed, rate=90.2k reads per second             ||
#> ||   36% completed, 3.1 mins elapsed, rate=93.7k reads per second             ||
#> ||   39% completed, 3.3 mins elapsed, rate=60.7k reads per second             ||
#> ||   40% completed, 3.3 mins elapsed, rate=60.9k reads per second             ||
#> ||   41% completed, 3.4 mins elapsed, rate=61.2k reads per second             ||
#> ||   41% completed, 3.4 mins elapsed, rate=61.4k reads per second             ||
#> ||   42% completed, 3.5 mins elapsed, rate=61.6k reads per second             ||
#> ||   43% completed, 3.5 mins elapsed, rate=61.8k reads per second             ||
#> ||   44% completed, 3.6 mins elapsed, rate=62.0k reads per second             ||
#> ||   44% completed, 3.6 mins elapsed, rate=62.2k reads per second             ||
#> ||   45% completed, 3.7 mins elapsed, rate=62.4k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   46% completed, 3.8 mins elapsed, rate=87.0k reads per second             ||
#> ||   53% completed, 4.1 mins elapsed, rate=89.6k reads per second             ||
#> ||   59% completed, 4.4 mins elapsed, rate=91.7k reads per second             ||
#> ||   62% completed, 4.6 mins elapsed, rate=68.8k reads per second             ||
#> ||   63% completed, 4.6 mins elapsed, rate=68.9k reads per second             ||
#> ||   64% completed, 4.7 mins elapsed, rate=69.0k reads per second             ||
#> ||   65% completed, 4.7 mins elapsed, rate=69.0k reads per second             ||
#> ||   65% completed, 4.8 mins elapsed, rate=69.1k reads per second             ||
#> ||   66% completed, 4.8 mins elapsed, rate=69.2k reads per second             ||
#> ||   67% completed, 4.9 mins elapsed, rate=69.2k reads per second             ||
#> ||   68% completed, 4.9 mins elapsed, rate=69.3k reads per second             ||
#> ||   69% completed, 5.0 mins elapsed, rate=69.3k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   69% completed, 5.1 mins elapsed, rate=87.5k reads per second             ||
#> ||   76% completed, 5.4 mins elapsed, rate=89.2k reads per second             ||
#> ||   83% completed, 5.7 mins elapsed, rate=90.8k reads per second             ||
#> ||   86% completed, 5.9 mins elapsed, rate=73.2k reads per second             ||
#> ||   86% completed, 5.9 mins elapsed, rate=73.2k reads per second             ||
#> ||   87% completed, 6.0 mins elapsed, rate=73.2k reads per second             ||
#> ||   88% completed, 6.0 mins elapsed, rate=73.2k reads per second             ||
#> ||   89% completed, 6.1 mins elapsed, rate=73.2k reads per second             ||
#> ||   90% completed, 6.1 mins elapsed, rate=73.2k reads per second             ||
#> ||   90% completed, 6.2 mins elapsed, rate=73.2k reads per second             ||
#> ||   91% completed, 6.3 mins elapsed, rate=73.3k reads per second             ||
#> ||   92% completed, 6.3 mins elapsed, rate=73.3k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   93% completed, 6.5 mins elapsed, rate=87.5k reads per second             ||
#> ||   97% completed, 6.7 mins elapsed, rate=73.1k reads per second             ||
#> ||   98% completed, 6.7 mins elapsed, rate=73.1k reads per second             ||
#> ||   98% completed, 6.7 mins elapsed, rate=73.1k reads per second             ||
#> ||   98% completed, 6.7 mins elapsed, rate=73.1k reads per second             ||
#> ||   98% completed, 6.8 mins elapsed, rate=73.1k reads per second             ||
#> ||   99% completed, 6.8 mins elapsed, rate=73.1k reads per second             ||
#> ||   99% completed, 6.8 mins elapsed, rate=73.1k reads per second             ||
#> ||   99% completed, 6.8 mins elapsed, rate=73.1k reads per second             ||
#> ||   99% completed, 6.8 mins elapsed, rate=73.1k reads per second             ||
#> ||                                                                            ||
#> ||                           Completed successfully.                          ||
#> ||                                                                            ||
#> \\====================================    ====================================//
#> 
#> //================================   Summary =================================\\
#> ||                                                                            ||
#> ||                 Total reads : 30,000,000                                   ||
#> ||                      Mapped : 21,351,123 (71.2%)                           ||
#> ||             Uniquely mapped : 21,351,123                                   ||
#> ||               Multi-mapping : 0                                            ||
#> ||                                                                            ||
#> ||                    Unmapped : 8,648,877                                    ||
#> ||                                                                            ||
#> ||                      Indels : 0                                            ||
#> ||                                                                            ||
#> ||                     WARNING : Phred offset (33) incorrect?                 ||
#> ||                                                                            ||
#> ||                Running time : 7.5 minutes                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> [1] "Index directory already exists remove it if you want to recreate it"
#> 
#>         ==========     _____ _    _ ____  _____  ______          _____  
#>         =====         / ____| |  | |  _ \|  __ \|  ____|   /\   |  __ \ 
#>           =====      | (___ | |  | | |_) | |__) | |__     /  \  | |  | |
#>             ====      \___ \| |  | |  _ <|  _  /|  __|   / /\ \ | |  | |
#>               ====    ____) | |__| | |_) | | \ \| |____ / ____ \| |__| |
#>         ==========   |_____/ \____/|____/|_|  \_\______/_/    \_\_____/
#>        Rsubread 2.20.0
#> 
#> //================================= setting ==================================\\
#> ||                                                                            ||
#> || Function      : Read alignment (DNA-Seq)                                   ||
#> || Input file    : prepro.PPP-BG.15M_CA_valid.fastq.gz                        ||
#> || Output file   : prepro.PPP-BG.15M_CA_valid.fastq.gz.bam (BAM), Sorted      ||
#> || Index name    : index                                                      ||
#> ||                                                                            ||
#> ||                    ------------------------------------                    ||
#> ||                                                                            ||
#> ||                               Threads : 1                                  ||
#> ||                          Phred offset : 33                                 ||
#> ||                             Min votes : 3 / 10                             ||
#> ||                        Max mismatches : 1                                  ||
#> ||                      Max indel length : 0                                  ||
#> ||            Report multi-mapping reads : no                                 ||
#> || Max alignments per multi-mapping read : 1                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> //================ Running (01-Nov-2025 18:59:22, pid=392792) ================\\
#> ||                                                                            ||
#> || Check the input reads.                                                     ||
#> || The input file contains base space reads.                                  ||
#> || Initialise the memory objects.                                             ||
#> || Estimate the mean read length.                                             ||
#> || WARNING  - The specified Phred score offset (33) seems incorrect.          ||
#> ||            ASCII values of the quality scores of read bases included in    ||
#> ||            the first 3000 reads were found to be within the range of       ||
#> ||            [63,63].                                                        ||
#> ||                                                                            ||
#> || Create the output BAM file.                                                ||
#> || Check the index.                                                           ||
#> || Init the voting space.                                                     ||
#> || Global environment is initialised.                                         ||
#> || Load the 1-th index block...                                               ||
#> || The index block has been loaded.                                           ||
#> || Start read mapping in chunk.                                               ||
#> ||    0% completed, 1.1 mins elapsed, rate=77.6k reads per second             ||
#> ||    6% completed, 1.4 mins elapsed, rate=115.3k reads per second            ||
#> ||   13% completed, 1.7 mins elapsed, rate=115.2k reads per second            ||
#> ||   16% completed, 1.9 mins elapsed, rate=43.2k reads per second             ||
#> ||   17% completed, 2.0 mins elapsed, rate=43.7k reads per second             ||
#> ||   17% completed, 2.0 mins elapsed, rate=44.3k reads per second             ||
#> ||   18% completed, 2.1 mins elapsed, rate=44.8k reads per second             ||
#> ||   19% completed, 2.1 mins elapsed, rate=45.3k reads per second             ||
#> ||   20% completed, 2.2 mins elapsed, rate=45.8k reads per second             ||
#> ||   20% completed, 2.3 mins elapsed, rate=46.2k reads per second             ||
#> ||   21% completed, 2.3 mins elapsed, rate=46.7k reads per second             ||
#> ||   22% completed, 2.4 mins elapsed, rate=47.1k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   23% completed, 2.5 mins elapsed, rate=83.6k reads per second             ||
#> ||   29% completed, 2.8 mins elapsed, rate=89.5k reads per second             ||
#> ||   36% completed, 3.1 mins elapsed, rate=93.8k reads per second             ||
#> ||   39% completed, 3.3 mins elapsed, rate=60.8k reads per second             ||
#> ||   40% completed, 3.3 mins elapsed, rate=60.8k reads per second             ||
#> ||   41% completed, 3.4 mins elapsed, rate=60.9k reads per second             ||
#> ||   41% completed, 3.4 mins elapsed, rate=60.9k reads per second             ||
#> ||   42% completed, 3.5 mins elapsed, rate=61.0k reads per second             ||
#> ||   43% completed, 3.6 mins elapsed, rate=61.0k reads per second             ||
#> ||   44% completed, 3.6 mins elapsed, rate=61.0k reads per second             ||
#> ||   45% completed, 3.7 mins elapsed, rate=61.0k reads per second             ||
#> ||   45% completed, 3.8 mins elapsed, rate=61.1k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   46% completed, 3.9 mins elapsed, rate=84.5k reads per second             ||
#> ||   53% completed, 4.2 mins elapsed, rate=87.7k reads per second             ||
#> ||   59% completed, 4.5 mins elapsed, rate=90.3k reads per second             ||
#> ||   62% completed, 4.6 mins elapsed, rate=68.1k reads per second             ||
#> ||   63% completed, 4.7 mins elapsed, rate=68.0k reads per second             ||
#> ||   64% completed, 4.7 mins elapsed, rate=68.0k reads per second             ||
#> ||   65% completed, 4.8 mins elapsed, rate=67.9k reads per second             ||
#> ||   66% completed, 4.9 mins elapsed, rate=67.8k reads per second             ||
#> ||   66% completed, 4.9 mins elapsed, rate=67.8k reads per second             ||
#> ||   67% completed, 5.0 mins elapsed, rate=67.7k reads per second             ||
#> ||   68% completed, 5.1 mins elapsed, rate=67.7k reads per second             ||
#> ||   69% completed, 5.1 mins elapsed, rate=67.6k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   69% completed, 5.3 mins elapsed, rate=85.1k reads per second             ||
#> ||   76% completed, 5.5 mins elapsed, rate=87.2k reads per second             ||
#> ||   83% completed, 5.8 mins elapsed, rate=89.1k reads per second             ||
#> ||   86% completed, 6.0 mins elapsed, rate=72.2k reads per second             ||
#> ||   86% completed, 6.0 mins elapsed, rate=72.1k reads per second             ||
#> ||   87% completed, 6.1 mins elapsed, rate=72.0k reads per second             ||
#> ||   88% completed, 6.2 mins elapsed, rate=71.9k reads per second             ||
#> ||   89% completed, 6.2 mins elapsed, rate=71.8k reads per second             ||
#> ||   90% completed, 6.3 mins elapsed, rate=71.7k reads per second             ||
#> ||   90% completed, 6.3 mins elapsed, rate=71.7k reads per second             ||
#> ||   91% completed, 6.4 mins elapsed, rate=71.6k reads per second             ||
#> ||   92% completed, 6.5 mins elapsed, rate=71.5k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   93% completed, 6.6 mins elapsed, rate=85.3k reads per second             ||
#> ||   97% completed, 6.8 mins elapsed, rate=71.8k reads per second             ||
#> ||   98% completed, 6.8 mins elapsed, rate=71.7k reads per second             ||
#> ||   98% completed, 6.9 mins elapsed, rate=71.7k reads per second             ||
#> ||   98% completed, 6.9 mins elapsed, rate=71.7k reads per second             ||
#> ||   98% completed, 6.9 mins elapsed, rate=71.7k reads per second             ||
#> ||   99% completed, 6.9 mins elapsed, rate=71.7k reads per second             ||
#> ||   99% completed, 6.9 mins elapsed, rate=71.6k reads per second             ||
#> ||   99% completed, 7.0 mins elapsed, rate=71.6k reads per second             ||
#> ||   99% completed, 7.0 mins elapsed, rate=71.6k reads per second             ||
#> ||                                                                            ||
#> ||                           Completed successfully.                          ||
#> ||                                                                            ||
#> \\====================================    ====================================//
#> 
#> //================================   Summary =================================\\
#> ||                                                                            ||
#> ||                 Total reads : 30,000,000                                   ||
#> ||                      Mapped : 5,623,606 (18.7%)                            ||
#> ||             Uniquely mapped : 5,623,606                                    ||
#> ||               Multi-mapping : 0                                            ||
#> ||                                                                            ||
#> ||                    Unmapped : 24,376,394                                   ||
#> ||                                                                            ||
#> ||                      Indels : 0                                            ||
#> ||                                                                            ||
#> ||                     WARNING : Phred offset (33) incorrect?                 ||
#> ||                                                                            ||
#> ||                Running time : 7.7 minutes                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> [1] "Index directory already exists remove it if you want to recreate it"
#> 
#>         ==========     _____ _    _ ____  _____  ______          _____  
#>         =====         / ____| |  | |  _ \|  __ \|  ____|   /\   |  __ \ 
#>           =====      | (___ | |  | | |_) | |__) | |__     /  \  | |  | |
#>             ====      \___ \| |  | |  _ <|  _  /|  __|   / /\ \ | |  | |
#>               ====    ____) | |__| | |_) | | \ \| |____ / ____ \| |__| |
#>         ==========   |_____/ \____/|____/|_|  \_\______/_/    \_\_____/
#>        Rsubread 2.20.0
#> 
#> //================================= setting ==================================\\
#> ||                                                                            ||
#> || Function      : Read alignment (DNA-Seq)                                   ||
#> || Input file    : prepro.PPP-BG.15M_CC_valid.fastq.gz                        ||
#> || Output file   : prepro.PPP-BG.15M_CC_valid.fastq.gz.bam (BAM), Sorted      ||
#> || Index name    : index                                                      ||
#> ||                                                                            ||
#> ||                    ------------------------------------                    ||
#> ||                                                                            ||
#> ||                               Threads : 1                                  ||
#> ||                          Phred offset : 33                                 ||
#> ||                             Min votes : 3 / 10                             ||
#> ||                        Max mismatches : 1                                  ||
#> ||                      Max indel length : 0                                  ||
#> ||            Report multi-mapping reads : no                                 ||
#> || Max alignments per multi-mapping read : 1                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> //================ Running (01-Nov-2025 19:07:04, pid=392792) ================\\
#> ||                                                                            ||
#> || Check the input reads.                                                     ||
#> || The input file contains base space reads.                                  ||
#> || Initialise the memory objects.                                             ||
#> || Estimate the mean read length.                                             ||
#> || WARNING  - The specified Phred score offset (33) seems incorrect.          ||
#> ||            ASCII values of the quality scores of read bases included in    ||
#> ||            the first 3000 reads were found to be within the range of       ||
#> ||            [63,63].                                                        ||
#> ||                                                                            ||
#> || Create the output BAM file.                                                ||
#> || Check the index.                                                           ||
#> || Init the voting space.                                                     ||
#> || Global environment is initialised.                                         ||
#> || Load the 1-th index block...                                               ||
#> || The index block has been loaded.                                           ||
#> || Start read mapping in chunk.                                               ||
#> ||    0% completed, 1.2 mins elapsed, rate=101.1k reads per second            ||
#> ||    6% completed, 1.4 mins elapsed, rate=115.2k reads per second            ||
#> ||   13% completed, 1.7 mins elapsed, rate=115.1k reads per second            ||
#> ||   16% completed, 1.9 mins elapsed, rate=43.0k reads per second             ||
#> ||   17% completed, 2.0 mins elapsed, rate=43.6k reads per second             ||
#> ||   17% completed, 2.0 mins elapsed, rate=44.2k reads per second             ||
#> ||   18% completed, 2.1 mins elapsed, rate=44.7k reads per second             ||
#> ||   19% completed, 2.1 mins elapsed, rate=45.2k reads per second             ||
#> ||   20% completed, 2.2 mins elapsed, rate=45.7k reads per second             ||
#> ||   20% completed, 2.3 mins elapsed, rate=46.2k reads per second             ||
#> ||   21% completed, 2.3 mins elapsed, rate=46.6k reads per second             ||
#> ||   22% completed, 2.4 mins elapsed, rate=47.0k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   23% completed, 2.6 mins elapsed, rate=83.0k reads per second             ||
#> ||   29% completed, 2.8 mins elapsed, rate=89.0k reads per second             ||
#> ||   36% completed, 3.1 mins elapsed, rate=93.3k reads per second             ||
#> ||   39% completed, 3.3 mins elapsed, rate=60.5k reads per second             ||
#> ||   40% completed, 3.3 mins elapsed, rate=60.6k reads per second             ||
#> ||   41% completed, 3.4 mins elapsed, rate=60.6k reads per second             ||
#> ||   41% completed, 3.5 mins elapsed, rate=60.7k reads per second             ||
#> ||   42% completed, 3.5 mins elapsed, rate=60.7k reads per second             ||
#> ||   43% completed, 3.6 mins elapsed, rate=60.7k reads per second             ||
#> ||   44% completed, 3.6 mins elapsed, rate=60.8k reads per second             ||
#> ||   45% completed, 3.7 mins elapsed, rate=60.8k reads per second             ||
#> ||   45% completed, 3.8 mins elapsed, rate=60.8k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   46% completed, 3.9 mins elapsed, rate=84.8k reads per second             ||
#> ||   53% completed, 4.2 mins elapsed, rate=88.0k reads per second             ||
#> ||   59% completed, 4.5 mins elapsed, rate=90.6k reads per second             ||
#> ||   62% completed, 4.6 mins elapsed, rate=68.1k reads per second             ||
#> ||   63% completed, 4.7 mins elapsed, rate=68.1k reads per second             ||
#> ||   64% completed, 4.7 mins elapsed, rate=68.0k reads per second             ||
#> ||   65% completed, 4.8 mins elapsed, rate=67.9k reads per second             ||
#> ||   65% completed, 4.9 mins elapsed, rate=67.9k reads per second             ||
#> ||   66% completed, 4.9 mins elapsed, rate=67.8k reads per second             ||
#> ||   67% completed, 5.0 mins elapsed, rate=67.7k reads per second             ||
#> ||   68% completed, 5.0 mins elapsed, rate=67.7k reads per second             ||
#> ||   69% completed, 5.1 mins elapsed, rate=67.6k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   69% completed, 5.2 mins elapsed, rate=85.4k reads per second             ||
#> ||   76% completed, 5.5 mins elapsed, rate=87.6k reads per second             ||
#> ||   83% completed, 5.8 mins elapsed, rate=89.4k reads per second             ||
#> ||   86% completed, 6.0 mins elapsed, rate=72.2k reads per second             ||
#> ||   86% completed, 6.0 mins elapsed, rate=72.1k reads per second             ||
#> ||   87% completed, 6.1 mins elapsed, rate=72.0k reads per second             ||
#> ||   88% completed, 6.2 mins elapsed, rate=72.0k reads per second             ||
#> ||   89% completed, 6.2 mins elapsed, rate=71.9k reads per second             ||
#> ||   90% completed, 6.3 mins elapsed, rate=71.8k reads per second             ||
#> ||   90% completed, 6.3 mins elapsed, rate=71.7k reads per second             ||
#> ||   91% completed, 6.4 mins elapsed, rate=71.6k reads per second             ||
#> ||   92% completed, 6.5 mins elapsed, rate=71.5k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   93% completed, 6.6 mins elapsed, rate=85.7k reads per second             ||
#> ||   97% completed, 6.8 mins elapsed, rate=72.0k reads per second             ||
#> ||   98% completed, 6.8 mins elapsed, rate=71.9k reads per second             ||
#> ||   98% completed, 6.8 mins elapsed, rate=71.9k reads per second             ||
#> ||   98% completed, 6.9 mins elapsed, rate=71.9k reads per second             ||
#> ||   98% completed, 6.9 mins elapsed, rate=71.9k reads per second             ||
#> ||   99% completed, 6.9 mins elapsed, rate=71.8k reads per second             ||
#> ||   99% completed, 6.9 mins elapsed, rate=71.8k reads per second             ||
#> ||   99% completed, 6.9 mins elapsed, rate=71.8k reads per second             ||
#> ||   99% completed, 7.0 mins elapsed, rate=71.8k reads per second             ||
#> ||                                                                            ||
#> ||                           Completed successfully.                          ||
#> ||                                                                            ||
#> \\====================================    ====================================//
#> 
#> //================================   Summary =================================\\
#> ||                                                                            ||
#> ||                 Total reads : 30,000,000                                   ||
#> ||                      Mapped : 5,627,855 (18.8%)                            ||
#> ||             Uniquely mapped : 5,627,855                                    ||
#> ||               Multi-mapping : 0                                            ||
#> ||                                                                            ||
#> ||                    Unmapped : 24,372,145                                   ||
#> ||                                                                            ||
#> ||                      Indels : 0                                            ||
#> ||                                                                            ||
#> ||                     WARNING : Phred offset (33) incorrect?                 ||
#> ||                                                                            ||
#> ||                Running time : 7.7 minutes                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> [1] "Index directory already exists remove it if you want to recreate it"
#> 
#>         ==========     _____ _    _ ____  _____  ______          _____  
#>         =====         / ____| |  | |  _ \|  __ \|  ____|   /\   |  __ \ 
#>           =====      | (___ | |  | | |_) | |__) | |__     /  \  | |  | |
#>             ====      \___ \| |  | |  _ <|  _  /|  __|   / /\ \ | |  | |
#>               ====    ____) | |__| | |_) | | \ \| |____ / ____ \| |__| |
#>         ==========   |_____/ \____/|____/|_|  \_\______/_/    \_\_____/
#>        Rsubread 2.20.0
#> 
#> //================================= setting ==================================\\
#> ||                                                                            ||
#> || Function      : Read alignment (DNA-Seq)                                   ||
#> || Input file    : prepro.PPP-BG.15M_CT_valid.fastq.gz                        ||
#> || Output file   : prepro.PPP-BG.15M_CT_valid.fastq.gz.bam (BAM), Sorted      ||
#> || Index name    : index                                                      ||
#> ||                                                                            ||
#> ||                    ------------------------------------                    ||
#> ||                                                                            ||
#> ||                               Threads : 1                                  ||
#> ||                          Phred offset : 33                                 ||
#> ||                             Min votes : 3 / 10                             ||
#> ||                        Max mismatches : 1                                  ||
#> ||                      Max indel length : 0                                  ||
#> ||            Report multi-mapping reads : no                                 ||
#> || Max alignments per multi-mapping read : 1                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> //================ Running (01-Nov-2025 19:14:45, pid=392792) ================\\
#> ||                                                                            ||
#> || Check the input reads.                                                     ||
#> || The input file contains base space reads.                                  ||
#> || Initialise the memory objects.                                             ||
#> || Estimate the mean read length.                                             ||
#> || WARNING  - The specified Phred score offset (33) seems incorrect.          ||
#> ||            ASCII values of the quality scores of read bases included in    ||
#> ||            the first 3000 reads were found to be within the range of       ||
#> ||            [63,63].                                                        ||
#> ||                                                                            ||
#> || Create the output BAM file.                                                ||
#> || Check the index.                                                           ||
#> || Init the voting space.                                                     ||
#> || Global environment is initialised.                                         ||
#> || Load the 1-th index block...                                               ||
#> || The index block has been loaded.                                           ||
#> || Start read mapping in chunk.                                               ||
#> ||    0% completed, 1.1 mins elapsed, rate=102.2k reads per second            ||
#> ||    6% completed, 1.4 mins elapsed, rate=114.9k reads per second            ||
#> ||   13% completed, 1.7 mins elapsed, rate=115.1k reads per second            ||
#> ||   16% completed, 1.9 mins elapsed, rate=43.3k reads per second             ||
#> ||   16% completed, 1.9 mins elapsed, rate=43.9k reads per second             ||
#> ||   17% completed, 2.0 mins elapsed, rate=44.5k reads per second             ||
#> ||   18% completed, 2.1 mins elapsed, rate=45.0k reads per second             ||
#> ||   19% completed, 2.1 mins elapsed, rate=45.5k reads per second             ||
#> ||   20% completed, 2.2 mins elapsed, rate=46.0k reads per second             ||
#> ||   20% completed, 2.3 mins elapsed, rate=46.4k reads per second             ||
#> ||   21% completed, 2.3 mins elapsed, rate=46.8k reads per second             ||
#> ||   22% completed, 2.4 mins elapsed, rate=47.3k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   23% completed, 2.5 mins elapsed, rate=83.9k reads per second             ||
#> ||   29% completed, 2.8 mins elapsed, rate=89.7k reads per second             ||
#> ||   36% completed, 3.1 mins elapsed, rate=94.0k reads per second             ||
#> ||   39% completed, 3.2 mins elapsed, rate=60.9k reads per second             ||
#> ||   40% completed, 3.3 mins elapsed, rate=60.9k reads per second             ||
#> ||   40% completed, 3.4 mins elapsed, rate=60.9k reads per second             ||
#> ||   41% completed, 3.4 mins elapsed, rate=61.0k reads per second             ||
#> ||   42% completed, 3.5 mins elapsed, rate=61.0k reads per second             ||
#> ||   43% completed, 3.6 mins elapsed, rate=61.1k reads per second             ||
#> ||   44% completed, 3.6 mins elapsed, rate=61.1k reads per second             ||
#> ||   44% completed, 3.7 mins elapsed, rate=61.2k reads per second             ||
#> ||   45% completed, 3.7 mins elapsed, rate=61.2k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   46% completed, 3.9 mins elapsed, rate=85.5k reads per second             ||
#> ||   53% completed, 4.2 mins elapsed, rate=88.6k reads per second             ||
#> ||   59% completed, 4.4 mins elapsed, rate=91.1k reads per second             ||
#> ||   62% completed, 4.6 mins elapsed, rate=68.5k reads per second             ||
#> ||   63% completed, 4.6 mins elapsed, rate=68.5k reads per second             ||
#> ||   64% completed, 4.7 mins elapsed, rate=68.4k reads per second             ||
#> ||   65% completed, 4.8 mins elapsed, rate=68.3k reads per second             ||
#> ||   65% completed, 4.8 mins elapsed, rate=68.3k reads per second             ||
#> ||   66% completed, 4.9 mins elapsed, rate=68.2k reads per second             ||
#> ||   67% completed, 5.0 mins elapsed, rate=68.2k reads per second             ||
#> ||   68% completed, 5.0 mins elapsed, rate=68.1k reads per second             ||
#> ||   68% completed, 5.1 mins elapsed, rate=68.0k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   69% completed, 5.2 mins elapsed, rate=86.0k reads per second             ||
#> ||   76% completed, 5.5 mins elapsed, rate=88.1k reads per second             ||
#> ||   83% completed, 5.8 mins elapsed, rate=90.0k reads per second             ||
#> ||   86% completed, 5.9 mins elapsed, rate=72.7k reads per second             ||
#> ||   86% completed, 6.0 mins elapsed, rate=72.6k reads per second             ||
#> ||   87% completed, 6.0 mins elapsed, rate=72.5k reads per second             ||
#> ||   88% completed, 6.1 mins elapsed, rate=72.5k reads per second             ||
#> ||   89% completed, 6.2 mins elapsed, rate=72.4k reads per second             ||
#> ||   89% completed, 6.2 mins elapsed, rate=72.3k reads per second             ||
#> ||   90% completed, 6.3 mins elapsed, rate=72.2k reads per second             ||
#> ||   91% completed, 6.4 mins elapsed, rate=72.1k reads per second             ||
#> ||   92% completed, 6.4 mins elapsed, rate=72.0k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   93% completed, 6.5 mins elapsed, rate=86.3k reads per second             ||
#> ||   97% completed, 6.8 mins elapsed, rate=72.5k reads per second             ||
#> ||   98% completed, 6.8 mins elapsed, rate=72.5k reads per second             ||
#> ||   98% completed, 6.8 mins elapsed, rate=72.5k reads per second             ||
#> ||   98% completed, 6.8 mins elapsed, rate=72.4k reads per second             ||
#> ||   98% completed, 6.8 mins elapsed, rate=72.4k reads per second             ||
#> ||   99% completed, 6.8 mins elapsed, rate=72.4k reads per second             ||
#> ||   99% completed, 6.9 mins elapsed, rate=72.4k reads per second             ||
#> ||   99% completed, 6.9 mins elapsed, rate=72.3k reads per second             ||
#> ||   99% completed, 6.9 mins elapsed, rate=72.3k reads per second             ||
#> ||                                                                            ||
#> ||                           Completed successfully.                          ||
#> ||                                                                            ||
#> \\====================================    ====================================//
#> 
#> //================================   Summary =================================\\
#> ||                                                                            ||
#> ||                 Total reads : 30,000,000                                   ||
#> ||                      Mapped : 5,625,823 (18.8%)                            ||
#> ||             Uniquely mapped : 5,625,823                                    ||
#> ||               Multi-mapping : 0                                            ||
#> ||                                                                            ||
#> ||                    Unmapped : 24,374,177                                   ||
#> ||                                                                            ||
#> ||                      Indels : 0                                            ||
#> ||                                                                            ||
#> ||                     WARNING : Phred offset (33) incorrect?                 ||
#> ||                                                                            ||
#> ||                Running time : 7.6 minutes                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> [1] "Index directory already exists remove it if you want to recreate it"
#> 
#>         ==========     _____ _    _ ____  _____  ______          _____  
#>         =====         / ____| |  | |  _ \|  __ \|  ____|   /\   |  __ \ 
#>           =====      | (___ | |  | | |_) | |__) | |__     /  \  | |  | |
#>             ====      \___ \| |  | |  _ <|  _  /|  __|   / /\ \ | |  | |
#>               ====    ____) | |__| | |_) | | \ \| |____ / ____ \| |__| |
#>         ==========   |_____/ \____/|____/|_|  \_\______/_/    \_\_____/
#>        Rsubread 2.20.0
#> 
#> //================================= setting ==================================\\
#> ||                                                                            ||
#> || Function      : Read alignment (DNA-Seq)                                   ||
#> || Input file    : prepro.PPP-BG.20M_AA_valid.fastq.gz                        ||
#> || Output file   : prepro.PPP-BG.20M_AA_valid.fastq.gz.bam (BAM), Sorted      ||
#> || Index name    : index                                                      ||
#> ||                                                                            ||
#> ||                    ------------------------------------                    ||
#> ||                                                                            ||
#> ||                               Threads : 1                                  ||
#> ||                          Phred offset : 33                                 ||
#> ||                             Min votes : 3 / 10                             ||
#> ||                        Max mismatches : 1                                  ||
#> ||                      Max indel length : 0                                  ||
#> ||            Report multi-mapping reads : no                                 ||
#> || Max alignments per multi-mapping read : 1                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> //================ Running (01-Nov-2025 19:22:27, pid=392792) ================\\
#> ||                                                                            ||
#> || Check the input reads.                                                     ||
#> || The input file contains base space reads.                                  ||
#> || Initialise the memory objects.                                             ||
#> || Estimate the mean read length.                                             ||
#> || WARNING  - The specified Phred score offset (33) seems incorrect.          ||
#> ||            ASCII values of the quality scores of read bases included in    ||
#> ||            the first 3000 reads were found to be within the range of       ||
#> ||            [63,63].                                                        ||
#> ||                                                                            ||
#> || Create the output BAM file.                                                ||
#> || Check the index.                                                           ||
#> || Init the voting space.                                                     ||
#> || Global environment is initialised.                                         ||
#> || Load the 1-th index block...                                               ||
#> || The index block has been loaded.                                           ||
#> || Start read mapping in chunk.                                               ||
#> ||    0% completed, 1.2 mins elapsed, rate=98.8k reads per second             ||
#> ||    6% completed, 1.5 mins elapsed, rate=108.9k reads per second            ||
#> ||   12% completed, 1.9 mins elapsed, rate=42.4k reads per second             ||
#> ||   12% completed, 2.0 mins elapsed, rate=43.2k reads per second             ||
#> ||   13% completed, 2.0 mins elapsed, rate=44.0k reads per second             ||
#> ||   13% completed, 2.1 mins elapsed, rate=44.7k reads per second             ||
#> ||   14% completed, 2.1 mins elapsed, rate=45.4k reads per second             ||
#> ||   15% completed, 2.2 mins elapsed, rate=46.1k reads per second             ||
#> ||   15% completed, 2.2 mins elapsed, rate=46.8k reads per second             ||
#> ||   16% completed, 2.3 mins elapsed, rate=47.4k reads per second             ||
#> ||   16% completed, 2.3 mins elapsed, rate=48.0k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   17% completed, 2.5 mins elapsed, rate=84.6k reads per second             ||
#> ||   24% completed, 2.9 mins elapsed, rate=90.9k reads per second             ||
#> ||   29% completed, 3.3 mins elapsed, rate=60.4k reads per second             ||
#> ||   30% completed, 3.3 mins elapsed, rate=60.7k reads per second             ||
#> ||   30% completed, 3.4 mins elapsed, rate=60.9k reads per second             ||
#> ||   31% completed, 3.4 mins elapsed, rate=61.1k reads per second             ||
#> ||   31% completed, 3.5 mins elapsed, rate=61.3k reads per second             ||
#> ||   32% completed, 3.5 mins elapsed, rate=61.5k reads per second             ||
#> ||   33% completed, 3.6 mins elapsed, rate=61.7k reads per second             ||
#> ||   33% completed, 3.6 mins elapsed, rate=61.9k reads per second             ||
#> ||   34% completed, 3.7 mins elapsed, rate=62.1k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   34% completed, 3.9 mins elapsed, rate=86.4k reads per second             ||
#> ||   41% completed, 4.3 mins elapsed, rate=89.8k reads per second             ||
#> ||   47% completed, 4.6 mins elapsed, rate=68.4k reads per second             ||
#> ||   47% completed, 4.6 mins elapsed, rate=68.5k reads per second             ||
#> ||   48% completed, 4.7 mins elapsed, rate=68.6k reads per second             ||
#> ||   48% completed, 4.8 mins elapsed, rate=68.7k reads per second             ||
#> ||   49% completed, 4.8 mins elapsed, rate=68.7k reads per second             ||
#> ||   49% completed, 4.9 mins elapsed, rate=68.8k reads per second             ||
#> ||   50% completed, 4.9 mins elapsed, rate=68.9k reads per second             ||
#> ||   51% completed, 5.0 mins elapsed, rate=68.9k reads per second             ||
#> ||   51% completed, 5.0 mins elapsed, rate=69.0k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   52% completed, 5.2 mins elapsed, rate=87.1k reads per second             ||
#> ||   58% completed, 5.6 mins elapsed, rate=89.3k reads per second             ||
#> ||   64% completed, 5.9 mins elapsed, rate=72.8k reads per second             ||
#> ||   65% completed, 6.0 mins elapsed, rate=72.9k reads per second             ||
#> ||   65% completed, 6.0 mins elapsed, rate=72.9k reads per second             ||
#> ||   66% completed, 6.1 mins elapsed, rate=72.9k reads per second             ||
#> ||   66% completed, 6.1 mins elapsed, rate=72.9k reads per second             ||
#> ||   67% completed, 6.2 mins elapsed, rate=73.0k reads per second             ||
#> ||   67% completed, 6.2 mins elapsed, rate=73.0k reads per second             ||
#> ||   68% completed, 6.3 mins elapsed, rate=73.0k reads per second             ||
#> ||   69% completed, 6.3 mins elapsed, rate=73.0k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   69% completed, 6.5 mins elapsed, rate=87.4k reads per second             ||
#> ||   76% completed, 6.9 mins elapsed, rate=89.1k reads per second             ||
#> ||   81% completed, 7.2 mins elapsed, rate=75.6k reads per second             ||
#> ||   82% completed, 7.3 mins elapsed, rate=75.6k reads per second             ||
#> ||   83% completed, 7.3 mins elapsed, rate=75.6k reads per second             ||
#> ||   83% completed, 7.4 mins elapsed, rate=75.6k reads per second             ||
#> ||   84% completed, 7.4 mins elapsed, rate=75.6k reads per second             ||
#> ||   84% completed, 7.5 mins elapsed, rate=75.6k reads per second             ||
#> ||   85% completed, 7.5 mins elapsed, rate=75.6k reads per second             ||
#> ||   86% completed, 7.6 mins elapsed, rate=75.6k reads per second             ||
#> ||   86% completed, 7.6 mins elapsed, rate=75.6k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   87% completed, 7.8 mins elapsed, rate=87.6k reads per second             ||
#> ||   94% completed, 8.2 mins elapsed, rate=89.0k reads per second             ||
#> ||   96% completed, 8.3 mins elapsed, rate=76.8k reads per second             ||
#> ||   96% completed, 8.4 mins elapsed, rate=76.8k reads per second             ||
#> ||   97% completed, 8.4 mins elapsed, rate=76.8k reads per second             ||
#> ||   97% completed, 8.5 mins elapsed, rate=76.8k reads per second             ||
#> ||   97% completed, 8.5 mins elapsed, rate=76.8k reads per second             ||
#> ||   98% completed, 8.5 mins elapsed, rate=76.8k reads per second             ||
#> ||   98% completed, 8.6 mins elapsed, rate=76.8k reads per second             ||
#> ||   99% completed, 8.6 mins elapsed, rate=76.8k reads per second             ||
#> ||   99% completed, 8.6 mins elapsed, rate=76.8k reads per second             ||
#> ||                                                                            ||
#> ||                           Completed successfully.                          ||
#> ||                                                                            ||
#> \\====================================    ====================================//
#> 
#> //================================   Summary =================================\\
#> ||                                                                            ||
#> ||                 Total reads : 39,999,979                                   ||
#> ||                      Mapped : 28,475,514 (71.2%)                           ||
#> ||             Uniquely mapped : 28,475,514                                   ||
#> ||               Multi-mapping : 0                                            ||
#> ||                                                                            ||
#> ||                    Unmapped : 11,524,465                                   ||
#> ||                                                                            ||
#> ||                      Indels : 0                                            ||
#> ||                                                                            ||
#> ||                     WARNING : Phred offset (33) incorrect?                 ||
#> ||                                                                            ||
#> ||                Running time : 9.7 minutes                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> [1] "Index directory already exists remove it if you want to recreate it"
#> 
#>         ==========     _____ _    _ ____  _____  ______          _____  
#>         =====         / ____| |  | |  _ \|  __ \|  ____|   /\   |  __ \ 
#>           =====      | (___ | |  | | |_) | |__) | |__     /  \  | |  | |
#>             ====      \___ \| |  | |  _ <|  _  /|  __|   / /\ \ | |  | |
#>               ====    ____) | |__| | |_) | | \ \| |____ / ____ \| |__| |
#>         ==========   |_____/ \____/|____/|_|  \_\______/_/    \_\_____/
#>        Rsubread 2.20.0
#> 
#> //================================= setting ==================================\\
#> ||                                                                            ||
#> || Function      : Read alignment (DNA-Seq)                                   ||
#> || Input file    : prepro.PPP-BG.20M_AC_valid.fastq.gz                        ||
#> || Output file   : prepro.PPP-BG.20M_AC_valid.fastq.gz.bam (BAM), Sorted      ||
#> || Index name    : index                                                      ||
#> ||                                                                            ||
#> ||                    ------------------------------------                    ||
#> ||                                                                            ||
#> ||                               Threads : 1                                  ||
#> ||                          Phred offset : 33                                 ||
#> ||                             Min votes : 3 / 10                             ||
#> ||                        Max mismatches : 1                                  ||
#> ||                      Max indel length : 0                                  ||
#> ||            Report multi-mapping reads : no                                 ||
#> || Max alignments per multi-mapping read : 1                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> //================ Running (01-Nov-2025 19:32:07, pid=392792) ================\\
#> ||                                                                            ||
#> || Check the input reads.                                                     ||
#> || The input file contains base space reads.                                  ||
#> || Initialise the memory objects.                                             ||
#> || Estimate the mean read length.                                             ||
#> || WARNING  - The specified Phred score offset (33) seems incorrect.          ||
#> ||            ASCII values of the quality scores of read bases included in    ||
#> ||            the first 3000 reads were found to be within the range of       ||
#> ||            [63,63].                                                        ||
#> ||                                                                            ||
#> || Create the output BAM file.                                                ||
#> || Check the index.                                                           ||
#> || Init the voting space.                                                     ||
#> || Global environment is initialised.                                         ||
#> || Load the 1-th index block...                                               ||
#> || The index block has been loaded.                                           ||
#> || Start read mapping in chunk.                                               ||
#> ||    0% completed, 1.2 mins elapsed, rate=98.1k reads per second             ||
#> ||    6% completed, 1.5 mins elapsed, rate=108.8k reads per second            ||
#> ||   12% completed, 1.9 mins elapsed, rate=42.5k reads per second             ||
#> ||   12% completed, 2.0 mins elapsed, rate=43.3k reads per second             ||
#> ||   13% completed, 2.0 mins elapsed, rate=44.1k reads per second             ||
#> ||   13% completed, 2.1 mins elapsed, rate=44.8k reads per second             ||
#> ||   14% completed, 2.1 mins elapsed, rate=45.5k reads per second             ||
#> ||   15% completed, 2.2 mins elapsed, rate=46.2k reads per second             ||
#> ||   15% completed, 2.2 mins elapsed, rate=46.8k reads per second             ||
#> ||   16% completed, 2.3 mins elapsed, rate=47.5k reads per second             ||
#> ||   16% completed, 2.3 mins elapsed, rate=48.0k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   17% completed, 2.5 mins elapsed, rate=84.7k reads per second             ||
#> ||   24% completed, 2.9 mins elapsed, rate=91.0k reads per second             ||
#> ||   29% completed, 3.3 mins elapsed, rate=60.5k reads per second             ||
#> ||   30% completed, 3.3 mins elapsed, rate=60.8k reads per second             ||
#> ||   30% completed, 3.4 mins elapsed, rate=61.0k reads per second             ||
#> ||   31% completed, 3.4 mins elapsed, rate=61.2k reads per second             ||
#> ||   31% completed, 3.5 mins elapsed, rate=61.4k reads per second             ||
#> ||   32% completed, 3.5 mins elapsed, rate=61.6k reads per second             ||
#> ||   33% completed, 3.6 mins elapsed, rate=61.8k reads per second             ||
#> ||   33% completed, 3.6 mins elapsed, rate=62.0k reads per second             ||
#> ||   34% completed, 3.7 mins elapsed, rate=62.2k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   34% completed, 3.9 mins elapsed, rate=85.9k reads per second             ||
#> ||   41% completed, 4.3 mins elapsed, rate=89.3k reads per second             ||
#> ||   47% completed, 4.6 mins elapsed, rate=68.2k reads per second             ||
#> ||   47% completed, 4.7 mins elapsed, rate=68.3k reads per second             ||
#> ||   48% completed, 4.7 mins elapsed, rate=68.4k reads per second             ||
#> ||   48% completed, 4.8 mins elapsed, rate=68.5k reads per second             ||
#> ||   49% completed, 4.8 mins elapsed, rate=68.6k reads per second             ||
#> ||   50% completed, 4.9 mins elapsed, rate=68.6k reads per second             ||
#> ||   50% completed, 4.9 mins elapsed, rate=68.7k reads per second             ||
#> ||   51% completed, 5.0 mins elapsed, rate=68.8k reads per second             ||
#> ||   51% completed, 5.0 mins elapsed, rate=68.8k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   52% completed, 5.2 mins elapsed, rate=86.6k reads per second             ||
#> ||   59% completed, 5.6 mins elapsed, rate=89.0k reads per second             ||
#> ||   64% completed, 5.9 mins elapsed, rate=72.7k reads per second             ||
#> ||   65% completed, 6.0 mins elapsed, rate=72.7k reads per second             ||
#> ||   65% completed, 6.0 mins elapsed, rate=72.7k reads per second             ||
#> ||   66% completed, 6.1 mins elapsed, rate=72.7k reads per second             ||
#> ||   66% completed, 6.1 mins elapsed, rate=72.7k reads per second             ||
#> ||   67% completed, 6.2 mins elapsed, rate=72.8k reads per second             ||
#> ||   68% completed, 6.2 mins elapsed, rate=72.8k reads per second             ||
#> ||   68% completed, 6.3 mins elapsed, rate=72.8k reads per second             ||
#> ||   69% completed, 6.3 mins elapsed, rate=72.8k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   69% completed, 6.5 mins elapsed, rate=86.7k reads per second             ||
#> ||   76% completed, 6.9 mins elapsed, rate=88.5k reads per second             ||
#> ||   82% completed, 7.3 mins elapsed, rate=75.3k reads per second             ||
#> ||   82% completed, 7.3 mins elapsed, rate=75.3k reads per second             ||
#> ||   83% completed, 7.4 mins elapsed, rate=75.3k reads per second             ||
#> ||   83% completed, 7.4 mins elapsed, rate=75.3k reads per second             ||
#> ||   84% completed, 7.5 mins elapsed, rate=75.3k reads per second             ||
#> ||   85% completed, 7.5 mins elapsed, rate=75.3k reads per second             ||
#> ||   85% completed, 7.6 mins elapsed, rate=75.3k reads per second             ||
#> ||   86% completed, 7.6 mins elapsed, rate=75.3k reads per second             ||
#> ||   86% completed, 7.7 mins elapsed, rate=75.3k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   87% completed, 7.8 mins elapsed, rate=87.1k reads per second             ||
#> ||   94% completed, 8.2 mins elapsed, rate=88.5k reads per second             ||
#> ||   96% completed, 8.4 mins elapsed, rate=76.5k reads per second             ||
#> ||   96% completed, 8.4 mins elapsed, rate=76.5k reads per second             ||
#> ||   97% completed, 8.5 mins elapsed, rate=76.5k reads per second             ||
#> ||   97% completed, 8.5 mins elapsed, rate=76.5k reads per second             ||
#> ||   97% completed, 8.5 mins elapsed, rate=76.5k reads per second             ||
#> ||   98% completed, 8.6 mins elapsed, rate=76.5k reads per second             ||
#> ||   98% completed, 8.6 mins elapsed, rate=76.5k reads per second             ||
#> ||   99% completed, 8.6 mins elapsed, rate=76.5k reads per second             ||
#> ||   99% completed, 8.7 mins elapsed, rate=76.5k reads per second             ||
#> ||                                                                            ||
#> ||                           Completed successfully.                          ||
#> ||                                                                            ||
#> \\====================================    ====================================//
#> 
#> //================================   Summary =================================\\
#> ||                                                                            ||
#> ||                 Total reads : 39,999,968                                   ||
#> ||                      Mapped : 28,470,177 (71.2%)                           ||
#> ||             Uniquely mapped : 28,470,177                                   ||
#> ||               Multi-mapping : 0                                            ||
#> ||                                                                            ||
#> ||                    Unmapped : 11,529,791                                   ||
#> ||                                                                            ||
#> ||                      Indels : 0                                            ||
#> ||                                                                            ||
#> ||                     WARNING : Phred offset (33) incorrect?                 ||
#> ||                                                                            ||
#> ||                Running time : 9.7 minutes                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> [1] "Index directory already exists remove it if you want to recreate it"
#> 
#>         ==========     _____ _    _ ____  _____  ______          _____  
#>         =====         / ____| |  | |  _ \|  __ \|  ____|   /\   |  __ \ 
#>           =====      | (___ | |  | | |_) | |__) | |__     /  \  | |  | |
#>             ====      \___ \| |  | |  _ <|  _  /|  __|   / /\ \ | |  | |
#>               ====    ____) | |__| | |_) | | \ \| |____ / ____ \| |__| |
#>         ==========   |_____/ \____/|____/|_|  \_\______/_/    \_\_____/
#>        Rsubread 2.20.0
#> 
#> //================================= setting ==================================\\
#> ||                                                                            ||
#> || Function      : Read alignment (DNA-Seq)                                   ||
#> || Input file    : prepro.PPP-BG.20M_AT_valid.fastq.gz                        ||
#> || Output file   : prepro.PPP-BG.20M_AT_valid.fastq.gz.bam (BAM), Sorted      ||
#> || Index name    : index                                                      ||
#> ||                                                                            ||
#> ||                    ------------------------------------                    ||
#> ||                                                                            ||
#> ||                               Threads : 1                                  ||
#> ||                          Phred offset : 33                                 ||
#> ||                             Min votes : 3 / 10                             ||
#> ||                        Max mismatches : 1                                  ||
#> ||                      Max indel length : 0                                  ||
#> ||            Report multi-mapping reads : no                                 ||
#> || Max alignments per multi-mapping read : 1                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> //================ Running (01-Nov-2025 19:41:48, pid=392792) ================\\
#> ||                                                                            ||
#> || Check the input reads.                                                     ||
#> || The input file contains base space reads.                                  ||
#> || Initialise the memory objects.                                             ||
#> || Estimate the mean read length.                                             ||
#> || WARNING  - The specified Phred score offset (33) seems incorrect.          ||
#> ||            ASCII values of the quality scores of read bases included in    ||
#> ||            the first 3000 reads were found to be within the range of       ||
#> ||            [63,63].                                                        ||
#> ||                                                                            ||
#> || Create the output BAM file.                                                ||
#> || Check the index.                                                           ||
#> || Init the voting space.                                                     ||
#> || Global environment is initialised.                                         ||
#> || Load the 1-th index block...                                               ||
#> || The index block has been loaded.                                           ||
#> || Start read mapping in chunk.                                               ||
#> ||    0% completed, 1.1 mins elapsed, rate=97.2k reads per second             ||
#> ||    6% completed, 1.5 mins elapsed, rate=109.1k reads per second            ||
#> ||   12% completed, 1.9 mins elapsed, rate=42.6k reads per second             ||
#> ||   12% completed, 2.0 mins elapsed, rate=43.4k reads per second             ||
#> ||   13% completed, 2.0 mins elapsed, rate=44.2k reads per second             ||
#> ||   13% completed, 2.1 mins elapsed, rate=45.0k reads per second             ||
#> ||   14% completed, 2.1 mins elapsed, rate=45.7k reads per second             ||
#> ||   15% completed, 2.2 mins elapsed, rate=46.3k reads per second             ||
#> ||   15% completed, 2.2 mins elapsed, rate=47.0k reads per second             ||
#> ||   16% completed, 2.3 mins elapsed, rate=47.6k reads per second             ||
#> ||   16% completed, 2.3 mins elapsed, rate=48.2k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   17% completed, 2.5 mins elapsed, rate=85.8k reads per second             ||
#> ||   24% completed, 2.9 mins elapsed, rate=91.9k reads per second             ||
#> ||   29% completed, 3.2 mins elapsed, rate=61.0k reads per second             ||
#> ||   30% completed, 3.3 mins elapsed, rate=61.2k reads per second             ||
#> ||   30% completed, 3.4 mins elapsed, rate=61.4k reads per second             ||
#> ||   31% completed, 3.4 mins elapsed, rate=61.6k reads per second             ||
#> ||   31% completed, 3.5 mins elapsed, rate=61.8k reads per second             ||
#> ||   32% completed, 3.5 mins elapsed, rate=62.0k reads per second             ||
#> ||   33% completed, 3.6 mins elapsed, rate=62.2k reads per second             ||
#> ||   33% completed, 3.6 mins elapsed, rate=62.4k reads per second             ||
#> ||   34% completed, 3.7 mins elapsed, rate=62.6k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   34% completed, 3.8 mins elapsed, rate=86.9k reads per second             ||
#> ||   41% completed, 4.2 mins elapsed, rate=90.3k reads per second             ||
#> ||   47% completed, 4.6 mins elapsed, rate=68.8k reads per second             ||
#> ||   47% completed, 4.6 mins elapsed, rate=68.9k reads per second             ||
#> ||   48% completed, 4.7 mins elapsed, rate=69.0k reads per second             ||
#> ||   48% completed, 4.7 mins elapsed, rate=69.1k reads per second             ||
#> ||   49% completed, 4.8 mins elapsed, rate=69.1k reads per second             ||
#> ||   50% completed, 4.8 mins elapsed, rate=69.2k reads per second             ||
#> ||   50% completed, 4.9 mins elapsed, rate=69.3k reads per second             ||
#> ||   51% completed, 4.9 mins elapsed, rate=69.3k reads per second             ||
#> ||   51% completed, 5.0 mins elapsed, rate=69.4k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   52% completed, 5.2 mins elapsed, rate=87.2k reads per second             ||
#> ||   59% completed, 5.6 mins elapsed, rate=89.4k reads per second             ||
#> ||   64% completed, 5.9 mins elapsed, rate=73.0k reads per second             ||
#> ||   65% completed, 6.0 mins elapsed, rate=73.1k reads per second             ||
#> ||   65% completed, 6.0 mins elapsed, rate=73.1k reads per second             ||
#> ||   66% completed, 6.1 mins elapsed, rate=73.1k reads per second             ||
#> ||   66% completed, 6.1 mins elapsed, rate=73.1k reads per second             ||
#> ||   67% completed, 6.2 mins elapsed, rate=73.1k reads per second             ||
#> ||   68% completed, 6.2 mins elapsed, rate=73.1k reads per second             ||
#> ||   68% completed, 6.3 mins elapsed, rate=73.1k reads per second             ||
#> ||   69% completed, 6.3 mins elapsed, rate=73.2k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   69% completed, 6.5 mins elapsed, rate=87.4k reads per second             ||
#> ||   76% completed, 6.9 mins elapsed, rate=89.1k reads per second             ||
#> ||   82% completed, 7.2 mins elapsed, rate=75.8k reads per second             ||
#> ||   82% completed, 7.3 mins elapsed, rate=75.8k reads per second             ||
#> ||   83% completed, 7.3 mins elapsed, rate=75.8k reads per second             ||
#> ||   83% completed, 7.4 mins elapsed, rate=75.8k reads per second             ||
#> ||   84% completed, 7.4 mins elapsed, rate=75.8k reads per second             ||
#> ||   85% completed, 7.5 mins elapsed, rate=75.7k reads per second             ||
#> ||   85% completed, 7.5 mins elapsed, rate=75.7k reads per second             ||
#> ||   86% completed, 7.6 mins elapsed, rate=75.7k reads per second             ||
#> ||   86% completed, 7.6 mins elapsed, rate=75.7k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   87% completed, 7.8 mins elapsed, rate=87.4k reads per second             ||
#> ||   94% completed, 8.2 mins elapsed, rate=88.8k reads per second             ||
#> ||   96% completed, 8.3 mins elapsed, rate=76.8k reads per second             ||
#> ||   96% completed, 8.4 mins elapsed, rate=76.8k reads per second             ||
#> ||   97% completed, 8.4 mins elapsed, rate=76.8k reads per second             ||
#> ||   97% completed, 8.5 mins elapsed, rate=76.8k reads per second             ||
#> ||   97% completed, 8.5 mins elapsed, rate=76.8k reads per second             ||
#> ||   98% completed, 8.5 mins elapsed, rate=76.8k reads per second             ||
#> ||   98% completed, 8.6 mins elapsed, rate=76.8k reads per second             ||
#> ||   99% completed, 8.6 mins elapsed, rate=76.8k reads per second             ||
#> ||   99% completed, 8.6 mins elapsed, rate=76.8k reads per second             ||
#> ||                                                                            ||
#> ||                           Completed successfully.                          ||
#> ||                                                                            ||
#> \\====================================    ====================================//
#> 
#> //================================   Summary =================================\\
#> ||                                                                            ||
#> ||                 Total reads : 39,999,973                                   ||
#> ||                      Mapped : 28,470,305 (71.2%)                           ||
#> ||             Uniquely mapped : 28,470,305                                   ||
#> ||               Multi-mapping : 0                                            ||
#> ||                                                                            ||
#> ||                    Unmapped : 11,529,668                                   ||
#> ||                                                                            ||
#> ||                      Indels : 0                                            ||
#> ||                                                                            ||
#> ||                     WARNING : Phred offset (33) incorrect?                 ||
#> ||                                                                            ||
#> ||                Running time : 9.6 minutes                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> [1] "Index directory already exists remove it if you want to recreate it"
#> 
#>         ==========     _____ _    _ ____  _____  ______          _____  
#>         =====         / ____| |  | |  _ \|  __ \|  ____|   /\   |  __ \ 
#>           =====      | (___ | |  | | |_) | |__) | |__     /  \  | |  | |
#>             ====      \___ \| |  | |  _ <|  _  /|  __|   / /\ \ | |  | |
#>               ====    ____) | |__| | |_) | | \ \| |____ / ____ \| |__| |
#>         ==========   |_____/ \____/|____/|_|  \_\______/_/    \_\_____/
#>        Rsubread 2.20.0
#> 
#> //================================= setting ==================================\\
#> ||                                                                            ||
#> || Function      : Read alignment (DNA-Seq)                                   ||
#> || Input file    : prepro.PPP-BG.20M_CA_valid.fastq.gz                        ||
#> || Output file   : prepro.PPP-BG.20M_CA_valid.fastq.gz.bam (BAM), Sorted      ||
#> || Index name    : index                                                      ||
#> ||                                                                            ||
#> ||                    ------------------------------------                    ||
#> ||                                                                            ||
#> ||                               Threads : 1                                  ||
#> ||                          Phred offset : 33                                 ||
#> ||                             Min votes : 3 / 10                             ||
#> ||                        Max mismatches : 1                                  ||
#> ||                      Max indel length : 0                                  ||
#> ||            Report multi-mapping reads : no                                 ||
#> || Max alignments per multi-mapping read : 1                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> //================ Running (01-Nov-2025 19:51:26, pid=392792) ================\\
#> ||                                                                            ||
#> || Check the input reads.                                                     ||
#> || The input file contains base space reads.                                  ||
#> || Initialise the memory objects.                                             ||
#> || Estimate the mean read length.                                             ||
#> || WARNING  - The specified Phred score offset (33) seems incorrect.          ||
#> ||            ASCII values of the quality scores of read bases included in    ||
#> ||            the first 3000 reads were found to be within the range of       ||
#> ||            [63,63].                                                        ||
#> ||                                                                            ||
#> || Create the output BAM file.                                                ||
#> || Check the index.                                                           ||
#> || Init the voting space.                                                     ||
#> || Global environment is initialised.                                         ||
#> || Load the 1-th index block...                                               ||
#> || The index block has been loaded.                                           ||
#> || Start read mapping in chunk.                                               ||
#> ||    0% completed, 1.2 mins elapsed, rate=100.2k reads per second            ||
#> ||    6% completed, 1.5 mins elapsed, rate=115.0k reads per second            ||
#> ||   12% completed, 1.9 mins elapsed, rate=43.1k reads per second             ||
#> ||   12% completed, 2.0 mins elapsed, rate=43.7k reads per second             ||
#> ||   13% completed, 2.0 mins elapsed, rate=44.3k reads per second             ||
#> ||   13% completed, 2.1 mins elapsed, rate=44.8k reads per second             ||
#> ||   14% completed, 2.1 mins elapsed, rate=45.3k reads per second             ||
#> ||   15% completed, 2.2 mins elapsed, rate=45.8k reads per second             ||
#> ||   15% completed, 2.3 mins elapsed, rate=46.3k reads per second             ||
#> ||   16% completed, 2.3 mins elapsed, rate=46.7k reads per second             ||
#> ||   16% completed, 2.4 mins elapsed, rate=47.1k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   17% completed, 2.5 mins elapsed, rate=84.4k reads per second             ||
#> ||   24% completed, 2.9 mins elapsed, rate=91.8k reads per second             ||
#> ||   29% completed, 3.2 mins elapsed, rate=60.9k reads per second             ||
#> ||   30% completed, 3.3 mins elapsed, rate=61.0k reads per second             ||
#> ||   30% completed, 3.4 mins elapsed, rate=61.0k reads per second             ||
#> ||   31% completed, 3.4 mins elapsed, rate=61.0k reads per second             ||
#> ||   31% completed, 3.5 mins elapsed, rate=61.1k reads per second             ||
#> ||   32% completed, 3.6 mins elapsed, rate=61.1k reads per second             ||
#> ||   33% completed, 3.6 mins elapsed, rate=61.1k reads per second             ||
#> ||   33% completed, 3.7 mins elapsed, rate=61.2k reads per second             ||
#> ||   34% completed, 3.7 mins elapsed, rate=61.2k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   34% completed, 3.9 mins elapsed, rate=85.4k reads per second             ||
#> ||   41% completed, 4.3 mins elapsed, rate=89.5k reads per second             ||
#> ||   47% completed, 4.6 mins elapsed, rate=68.5k reads per second             ||
#> ||   47% completed, 4.7 mins elapsed, rate=68.4k reads per second             ||
#> ||   48% completed, 4.7 mins elapsed, rate=68.3k reads per second             ||
#> ||   48% completed, 4.8 mins elapsed, rate=68.3k reads per second             ||
#> ||   49% completed, 4.8 mins elapsed, rate=68.2k reads per second             ||
#> ||   50% completed, 4.9 mins elapsed, rate=68.1k reads per second             ||
#> ||   50% completed, 5.0 mins elapsed, rate=68.1k reads per second             ||
#> ||   51% completed, 5.0 mins elapsed, rate=68.0k reads per second             ||
#> ||   51% completed, 5.1 mins elapsed, rate=68.0k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   52% completed, 5.2 mins elapsed, rate=85.9k reads per second             ||
#> ||   59% completed, 5.6 mins elapsed, rate=88.7k reads per second             ||
#> ||   64% completed, 5.9 mins elapsed, rate=72.7k reads per second             ||
#> ||   65% completed, 6.0 mins elapsed, rate=72.6k reads per second             ||
#> ||   65% completed, 6.1 mins elapsed, rate=72.5k reads per second             ||
#> ||   66% completed, 6.1 mins elapsed, rate=72.4k reads per second             ||
#> ||   66% completed, 6.2 mins elapsed, rate=72.3k reads per second             ||
#> ||   67% completed, 6.2 mins elapsed, rate=72.2k reads per second             ||
#> ||   68% completed, 6.3 mins elapsed, rate=72.1k reads per second             ||
#> ||   68% completed, 6.4 mins elapsed, rate=72.0k reads per second             ||
#> ||   69% completed, 6.4 mins elapsed, rate=71.9k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   69% completed, 6.6 mins elapsed, rate=86.1k reads per second             ||
#> ||   76% completed, 6.9 mins elapsed, rate=88.2k reads per second             ||
#> ||   82% completed, 7.3 mins elapsed, rate=75.2k reads per second             ||
#> ||   82% completed, 7.3 mins elapsed, rate=75.1k reads per second             ||
#> ||   83% completed, 7.4 mins elapsed, rate=75.0k reads per second             ||
#> ||   83% completed, 7.5 mins elapsed, rate=74.9k reads per second             ||
#> ||   84% completed, 7.5 mins elapsed, rate=74.8k reads per second             ||
#> ||   85% completed, 7.6 mins elapsed, rate=74.7k reads per second             ||
#> ||   85% completed, 7.7 mins elapsed, rate=74.6k reads per second             ||
#> ||   86% completed, 7.7 mins elapsed, rate=74.5k reads per second             ||
#> ||   86% completed, 7.8 mins elapsed, rate=74.4k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   87% completed, 7.9 mins elapsed, rate=86.1k reads per second             ||
#> ||   94% completed, 8.3 mins elapsed, rate=87.8k reads per second             ||
#> ||   96% completed, 8.4 mins elapsed, rate=76.0k reads per second             ||
#> ||   96% completed, 8.5 mins elapsed, rate=75.9k reads per second             ||
#> ||   97% completed, 8.5 mins elapsed, rate=75.9k reads per second             ||
#> ||   97% completed, 8.6 mins elapsed, rate=75.8k reads per second             ||
#> ||   97% completed, 8.6 mins elapsed, rate=75.7k reads per second             ||
#> ||   98% completed, 8.7 mins elapsed, rate=75.7k reads per second             ||
#> ||   98% completed, 8.7 mins elapsed, rate=75.6k reads per second             ||
#> ||   99% completed, 8.7 mins elapsed, rate=75.5k reads per second             ||
#> ||   99% completed, 8.8 mins elapsed, rate=75.5k reads per second             ||
#> ||                                                                            ||
#> ||                           Completed successfully.                          ||
#> ||                                                                            ||
#> \\====================================    ====================================//
#> 
#> //================================   Summary =================================\\
#> ||                                                                            ||
#> ||                 Total reads : 39,999,987                                   ||
#> ||                      Mapped : 7,495,804 (18.7%)                            ||
#> ||             Uniquely mapped : 7,495,804                                    ||
#> ||               Multi-mapping : 0                                            ||
#> ||                                                                            ||
#> ||                    Unmapped : 32,504,183                                   ||
#> ||                                                                            ||
#> ||                      Indels : 0                                            ||
#> ||                                                                            ||
#> ||                     WARNING : Phred offset (33) incorrect?                 ||
#> ||                                                                            ||
#> ||                Running time : 9.8 minutes                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> [1] "Index directory already exists remove it if you want to recreate it"
#> 
#>         ==========     _____ _    _ ____  _____  ______          _____  
#>         =====         / ____| |  | |  _ \|  __ \|  ____|   /\   |  __ \ 
#>           =====      | (___ | |  | | |_) | |__) | |__     /  \  | |  | |
#>             ====      \___ \| |  | |  _ <|  _  /|  __|   / /\ \ | |  | |
#>               ====    ____) | |__| | |_) | | \ \| |____ / ____ \| |__| |
#>         ==========   |_____/ \____/|____/|_|  \_\______/_/    \_\_____/
#>        Rsubread 2.20.0
#> 
#> //================================= setting ==================================\\
#> ||                                                                            ||
#> || Function      : Read alignment (DNA-Seq)                                   ||
#> || Input file    : prepro.PPP-BG.20M_CC_valid.fastq.gz                        ||
#> || Output file   : prepro.PPP-BG.20M_CC_valid.fastq.gz.bam (BAM), Sorted      ||
#> || Index name    : index                                                      ||
#> ||                                                                            ||
#> ||                    ------------------------------------                    ||
#> ||                                                                            ||
#> ||                               Threads : 1                                  ||
#> ||                          Phred offset : 33                                 ||
#> ||                             Min votes : 3 / 10                             ||
#> ||                        Max mismatches : 1                                  ||
#> ||                      Max indel length : 0                                  ||
#> ||            Report multi-mapping reads : no                                 ||
#> || Max alignments per multi-mapping read : 1                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> //================ Running (01-Nov-2025 20:01:15, pid=392792) ================\\
#> ||                                                                            ||
#> || Check the input reads.                                                     ||
#> || The input file contains base space reads.                                  ||
#> || Initialise the memory objects.                                             ||
#> || Estimate the mean read length.                                             ||
#> || WARNING  - The specified Phred score offset (33) seems incorrect.          ||
#> ||            ASCII values of the quality scores of read bases included in    ||
#> ||            the first 3000 reads were found to be within the range of       ||
#> ||            [63,63].                                                        ||
#> ||                                                                            ||
#> || Create the output BAM file.                                                ||
#> || Check the index.                                                           ||
#> || Init the voting space.                                                     ||
#> || Global environment is initialised.                                         ||
#> || Load the 1-th index block...                                               ||
#> || The index block has been loaded.                                           ||
#> || Start read mapping in chunk.                                               ||
#> ||    0% completed, 1.1 mins elapsed, rate=99.1k reads per second             ||
#> ||    6% completed, 1.5 mins elapsed, rate=115.2k reads per second            ||
#> ||   12% completed, 1.9 mins elapsed, rate=43.2k reads per second             ||
#> ||   12% completed, 1.9 mins elapsed, rate=43.8k reads per second             ||
#> ||   13% completed, 2.0 mins elapsed, rate=44.4k reads per second             ||
#> ||   13% completed, 2.1 mins elapsed, rate=44.9k reads per second             ||
#> ||   14% completed, 2.1 mins elapsed, rate=45.4k reads per second             ||
#> ||   15% completed, 2.2 mins elapsed, rate=45.9k reads per second             ||
#> ||   15% completed, 2.3 mins elapsed, rate=46.3k reads per second             ||
#> ||   16% completed, 2.3 mins elapsed, rate=46.8k reads per second             ||
#> ||   16% completed, 2.4 mins elapsed, rate=47.2k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   17% completed, 2.5 mins elapsed, rate=84.0k reads per second             ||
#> ||   24% completed, 2.9 mins elapsed, rate=91.5k reads per second             ||
#> ||   29% completed, 3.2 mins elapsed, rate=61.0k reads per second             ||
#> ||   30% completed, 3.3 mins elapsed, rate=61.0k reads per second             ||
#> ||   30% completed, 3.4 mins elapsed, rate=61.0k reads per second             ||
#> ||   31% completed, 3.4 mins elapsed, rate=61.1k reads per second             ||
#> ||   31% completed, 3.5 mins elapsed, rate=61.1k reads per second             ||
#> ||   32% completed, 3.6 mins elapsed, rate=61.2k reads per second             ||
#> ||   33% completed, 3.6 mins elapsed, rate=61.2k reads per second             ||
#> ||   33% completed, 3.7 mins elapsed, rate=61.3k reads per second             ||
#> ||   34% completed, 3.7 mins elapsed, rate=61.3k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   34% completed, 3.9 mins elapsed, rate=85.2k reads per second             ||
#> ||   41% completed, 4.3 mins elapsed, rate=89.3k reads per second             ||
#> ||   47% completed, 4.6 mins elapsed, rate=68.4k reads per second             ||
#> ||   47% completed, 4.7 mins elapsed, rate=68.4k reads per second             ||
#> ||   48% completed, 4.7 mins elapsed, rate=68.3k reads per second             ||
#> ||   48% completed, 4.8 mins elapsed, rate=68.2k reads per second             ||
#> ||   49% completed, 4.8 mins elapsed, rate=68.1k reads per second             ||
#> ||   49% completed, 4.9 mins elapsed, rate=68.1k reads per second             ||
#> ||   50% completed, 5.0 mins elapsed, rate=68.0k reads per second             ||
#> ||   51% completed, 5.0 mins elapsed, rate=68.0k reads per second             ||
#> ||   51% completed, 5.1 mins elapsed, rate=67.9k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   52% completed, 5.2 mins elapsed, rate=85.6k reads per second             ||
#> ||   58% completed, 5.6 mins elapsed, rate=88.4k reads per second             ||
#> ||   64% completed, 6.0 mins elapsed, rate=72.4k reads per second             ||
#> ||   65% completed, 6.0 mins elapsed, rate=72.3k reads per second             ||
#> ||   65% completed, 6.1 mins elapsed, rate=72.2k reads per second             ||
#> ||   66% completed, 6.1 mins elapsed, rate=72.2k reads per second             ||
#> ||   66% completed, 6.2 mins elapsed, rate=72.1k reads per second             ||
#> ||   67% completed, 6.3 mins elapsed, rate=72.0k reads per second             ||
#> ||   67% completed, 6.3 mins elapsed, rate=71.9k reads per second             ||
#> ||   68% completed, 6.4 mins elapsed, rate=71.8k reads per second             ||
#> ||   69% completed, 6.4 mins elapsed, rate=71.7k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   69% completed, 6.6 mins elapsed, rate=85.7k reads per second             ||
#> ||   76% completed, 7.0 mins elapsed, rate=87.7k reads per second             ||
#> ||   81% completed, 7.3 mins elapsed, rate=74.9k reads per second             ||
#> ||   82% completed, 7.4 mins elapsed, rate=74.8k reads per second             ||
#> ||   83% completed, 7.4 mins elapsed, rate=74.7k reads per second             ||
#> ||   83% completed, 7.5 mins elapsed, rate=74.6k reads per second             ||
#> ||   84% completed, 7.6 mins elapsed, rate=74.5k reads per second             ||
#> ||   84% completed, 7.6 mins elapsed, rate=74.4k reads per second             ||
#> ||   85% completed, 7.7 mins elapsed, rate=74.3k reads per second             ||
#> ||   86% completed, 7.7 mins elapsed, rate=74.2k reads per second             ||
#> ||   86% completed, 7.8 mins elapsed, rate=74.1k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   87% completed, 7.9 mins elapsed, rate=85.7k reads per second             ||
#> ||   94% completed, 8.3 mins elapsed, rate=87.4k reads per second             ||
#> ||   96% completed, 8.5 mins elapsed, rate=75.8k reads per second             ||
#> ||   96% completed, 8.5 mins elapsed, rate=75.7k reads per second             ||
#> ||   97% completed, 8.6 mins elapsed, rate=75.6k reads per second             ||
#> ||   97% completed, 8.6 mins elapsed, rate=75.6k reads per second             ||
#> ||   97% completed, 8.6 mins elapsed, rate=75.5k reads per second             ||
#> ||   98% completed, 8.7 mins elapsed, rate=75.5k reads per second             ||
#> ||   98% completed, 8.7 mins elapsed, rate=75.4k reads per second             ||
#> ||   99% completed, 8.8 mins elapsed, rate=75.3k reads per second             ||
#> ||   99% completed, 8.8 mins elapsed, rate=75.3k reads per second             ||
#> ||                                                                            ||
#> ||                           Completed successfully.                          ||
#> ||                                                                            ||
#> \\====================================    ====================================//
#> 
#> //================================   Summary =================================\\
#> ||                                                                            ||
#> ||                 Total reads : 39,999,981                                   ||
#> ||                      Mapped : 7,502,630 (18.8%)                            ||
#> ||             Uniquely mapped : 7,502,630                                    ||
#> ||               Multi-mapping : 0                                            ||
#> ||                                                                            ||
#> ||                    Unmapped : 32,497,351                                   ||
#> ||                                                                            ||
#> ||                      Indels : 0                                            ||
#> ||                                                                            ||
#> ||                     WARNING : Phred offset (33) incorrect?                 ||
#> ||                                                                            ||
#> ||                Running time : 9.8 minutes                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> [1] "Index directory already exists remove it if you want to recreate it"
#> 
#>         ==========     _____ _    _ ____  _____  ______          _____  
#>         =====         / ____| |  | |  _ \|  __ \|  ____|   /\   |  __ \ 
#>           =====      | (___ | |  | | |_) | |__) | |__     /  \  | |  | |
#>             ====      \___ \| |  | |  _ <|  _  /|  __|   / /\ \ | |  | |
#>               ====    ____) | |__| | |_) | | \ \| |____ / ____ \| |__| |
#>         ==========   |_____/ \____/|____/|_|  \_\______/_/    \_\_____/
#>        Rsubread 2.20.0
#> 
#> //================================= setting ==================================\\
#> ||                                                                            ||
#> || Function      : Read alignment (DNA-Seq)                                   ||
#> || Input file    : prepro.PPP-BG.20M_CT_valid.fastq.gz                        ||
#> || Output file   : prepro.PPP-BG.20M_CT_valid.fastq.gz.bam (BAM), Sorted      ||
#> || Index name    : index                                                      ||
#> ||                                                                            ||
#> ||                    ------------------------------------                    ||
#> ||                                                                            ||
#> ||                               Threads : 1                                  ||
#> ||                          Phred offset : 33                                 ||
#> ||                             Min votes : 3 / 10                             ||
#> ||                        Max mismatches : 1                                  ||
#> ||                      Max indel length : 0                                  ||
#> ||            Report multi-mapping reads : no                                 ||
#> || Max alignments per multi-mapping read : 1                                  ||
#> ||                                                                            ||
#> \\============================================================================//
#> 
#> //================ Running (01-Nov-2025 20:11:06, pid=392792) ================\\
#> ||                                                                            ||
#> || Check the input reads.                                                     ||
#> || The input file contains base space reads.                                  ||
#> || Initialise the memory objects.                                             ||
#> || Estimate the mean read length.                                             ||
#> || WARNING  - The specified Phred score offset (33) seems incorrect.          ||
#> ||            ASCII values of the quality scores of read bases included in    ||
#> ||            the first 3000 reads were found to be within the range of       ||
#> ||            [63,63].                                                        ||
#> ||                                                                            ||
#> || Create the output BAM file.                                                ||
#> || Check the index.                                                           ||
#> || Init the voting space.                                                     ||
#> || Global environment is initialised.                                         ||
#> || Load the 1-th index block...                                               ||
#> || The index block has been loaded.                                           ||
#> || Start read mapping in chunk.                                               ||
#> ||    0% completed, 1.2 mins elapsed, rate=100.3k reads per second            ||
#> ||    6% completed, 1.5 mins elapsed, rate=116.0k reads per second            ||
#> ||   12% completed, 1.9 mins elapsed, rate=43.2k reads per second             ||
#> ||   12% completed, 2.0 mins elapsed, rate=43.8k reads per second             ||
#> ||   13% completed, 2.0 mins elapsed, rate=44.4k reads per second             ||
#> ||   13% completed, 2.1 mins elapsed, rate=44.9k reads per second             ||
#> ||   14% completed, 2.1 mins elapsed, rate=45.4k reads per second             ||
#> ||   15% completed, 2.2 mins elapsed, rate=45.8k reads per second             ||
#> ||   15% completed, 2.3 mins elapsed, rate=46.3k reads per second             ||
#> ||   16% completed, 2.3 mins elapsed, rate=46.7k reads per second             ||
#> ||   16% completed, 2.4 mins elapsed, rate=47.1k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   17% completed, 2.5 mins elapsed, rate=83.7k reads per second             ||
#> ||   24% completed, 2.9 mins elapsed, rate=91.1k reads per second             ||
#> ||   29% completed, 3.3 mins elapsed, rate=60.8k reads per second             ||
#> ||   30% completed, 3.3 mins elapsed, rate=60.8k reads per second             ||
#> ||   30% completed, 3.4 mins elapsed, rate=60.9k reads per second             ||
#> ||   31% completed, 3.4 mins elapsed, rate=60.9k reads per second             ||
#> ||   31% completed, 3.5 mins elapsed, rate=61.0k reads per second             ||
#> ||   32% completed, 3.6 mins elapsed, rate=61.0k reads per second             ||
#> ||   33% completed, 3.6 mins elapsed, rate=61.1k reads per second             ||
#> ||   33% completed, 3.7 mins elapsed, rate=61.1k reads per second             ||
#> ||   34% completed, 3.7 mins elapsed, rate=61.1k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   34% completed, 3.9 mins elapsed, rate=85.3k reads per second             ||
#> ||   41% completed, 4.3 mins elapsed, rate=89.4k reads per second             ||
#> ||   47% completed, 4.6 mins elapsed, rate=68.4k reads per second             ||
#> ||   47% completed, 4.7 mins elapsed, rate=68.4k reads per second             ||
#> ||   48% completed, 4.7 mins elapsed, rate=68.3k reads per second             ||
#> ||   48% completed, 4.8 mins elapsed, rate=68.2k reads per second             ||
#> ||   49% completed, 4.8 mins elapsed, rate=68.2k reads per second             ||
#> ||   49% completed, 4.9 mins elapsed, rate=68.1k reads per second             ||
#> ||   50% completed, 5.0 mins elapsed, rate=68.1k reads per second             ||
#> ||   51% completed, 5.0 mins elapsed, rate=68.0k reads per second             ||
#> ||   51% completed, 5.1 mins elapsed, rate=67.9k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   52% completed, 5.2 mins elapsed, rate=85.7k reads per second             ||
#> ||   58% completed, 5.6 mins elapsed, rate=88.5k reads per second             ||
#> ||   64% completed, 5.9 mins elapsed, rate=72.5k reads per second             ||
#> ||   65% completed, 6.0 mins elapsed, rate=72.4k reads per second             ||
#> ||   65% completed, 6.1 mins elapsed, rate=72.3k reads per second             ||
#> ||   66% completed, 6.1 mins elapsed, rate=72.2k reads per second             ||
#> ||   66% completed, 6.2 mins elapsed, rate=72.2k reads per second             ||
#> ||   67% completed, 6.2 mins elapsed, rate=72.1k reads per second             ||
#> ||   67% completed, 6.3 mins elapsed, rate=72.0k reads per second             ||
#> ||   68% completed, 6.4 mins elapsed, rate=71.9k reads per second             ||
#> ||   69% completed, 6.4 mins elapsed, rate=71.9k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   69% completed, 6.6 mins elapsed, rate=86.0k reads per second             ||
#> ||   76% completed, 6.9 mins elapsed, rate=88.1k reads per second             ||
#> ||   82% completed, 7.3 mins elapsed, rate=75.1k reads per second             ||
#> ||   82% completed, 7.3 mins elapsed, rate=75.0k reads per second             ||
#> ||   83% completed, 7.4 mins elapsed, rate=74.9k reads per second             ||
#> ||   83% completed, 7.5 mins elapsed, rate=74.8k reads per second             ||
#> ||   84% completed, 7.5 mins elapsed, rate=74.7k reads per second             ||
#> ||   84% completed, 7.6 mins elapsed, rate=74.6k reads per second             ||
#> ||   85% completed, 7.7 mins elapsed, rate=74.5k reads per second             ||
#> ||   86% completed, 7.7 mins elapsed, rate=74.4k reads per second             ||
#> ||   86% completed, 7.8 mins elapsed, rate=74.4k reads per second             ||
#> || Start read mapping in chunk.                                               ||
#> ||   87% completed, 7.9 mins elapsed, rate=86.0k reads per second             ||
#> ||   94% completed, 8.3 mins elapsed, rate=87.8k reads per second             ||
#> ||   96% completed, 8.4 mins elapsed, rate=76.0k reads per second             ||
#> ||   96% completed, 8.5 mins elapsed, rate=75.9k reads per second             ||
#> ||   97% completed, 8.5 mins elapsed, rate=75.9k reads per second             ||
#> ||   97% completed, 8.6 mins elapsed, rate=75.8k reads per second             ||
#> ||   97% completed, 8.6 mins elapsed, rate=75.7k reads per second             ||
#> ||   98% completed, 8.7 mins elapsed, rate=75.7k reads per second             ||
#> ||   98% completed, 8.7 mins elapsed, rate=75.6k reads per second             ||
#> ||   99% completed, 8.8 mins elapsed, rate=75.6k reads per second             ||
#> ||   99% completed, 8.8 mins elapsed, rate=75.5k reads per second             ||
#> ||                                                                            ||
#> ||                           Completed successfully.                          ||
#> ||                                                                            ||
#> \\====================================    ====================================//
#> 
#> //================================   Summary =================================\\
#> ||                                                                            ||
#> ||                 Total reads : 39,999,991                                   ||
#> ||                      Mapped : 7,503,456 (18.8%)                            ||
#> ||             Uniquely mapped : 7,503,456                                    ||
#> ||               Multi-mapping : 0                                            ||
#> ||                                                                            ||
#> ||                    Unmapped : 32,496,535                                   ||
#> ||                                                                            ||
#> ||                      Indels : 0                                            ||
#> ||                                                                            ||
#> ||                     WARNING : Phred offset (33) incorrect?                 ||
#> ||                                                                            ||
#> ||                Running time : 9.8 minutes                                  ||
#> ||                                                                            ||
#> \\============================================================================//


prw_sr <- prw_sr %>%
  t %>%
  as_tibble() %>%
  select(time=Elapsed_Time_sec, 
         peak_ram=Peak_RAM_Used_MiB) %>%
  mutate(time=duration(as.numeric(time), units="seconds"), 
         peak_ram = as.numeric(peak_ram),
  nb_reads = c(12, 60, 120, 180, 240),
         aligner = "subread")
prw_sr %>%
  knitr::kable()

time

peak_ram

nb_reads

features

573.282000000007s (~9.55 minutes)

0.6

12

subread

1257.19799999999s (~20.95 minutes)

0.4

60

subread

1982.12s (~33.04 minutes)

0.4

120

subread

2751.33399999999s (~45.86 minutes)

0.4

180

subread

3507.849s (~58.46 minutes)

0.4

240

subread

Count table computation: quantify_from_bam_files

Measure time and maximum RAM in use for the quantify_from_bam_files function:

samples <-  c("PPP.r1", "PPP.r2", "PPP.r3", "BG.r1",   "BG.r2",  "BG.r3")

prw_counts <- sapply(1:length(dirout), function(i) {
  bam_files <- list.files(
    paste0(
      list.files(  
        dirout[i],
        pattern = "_valid.fastq.gz$", 
        full.names = TRUE
        ),
      "_MW2.fasta_mm1_bowtie"
    ),
    pattern=".bam$",
    full.names = TRUE
  )
  peakRAM(
    count_table <- quantify_from_bam_files(
      bam_files = bam_files,
      sample_ids = samples,
      fun = quantify_5prime,
      force = TRUE
    ))
  }
)
prw_counts <- prw_counts %>%
  t %>%
  as_tibble() %>%
  select(time=Elapsed_Time_sec, 
         peak_ram=Peak_RAM_Used_MiB) %>%
  mutate(time=duration(as.numeric(time), units="seconds"), 
         peak_ram = as.numeric(peak_ram),
         nb_reads = c(12, 60, 120, 180, 240),
         quantif = "quantify_5prime")
prw_counts %>%
  knitr::kable()

time

peak_ram

nb_reads

features

71.3050000000076s (~1.19 minutes)

476

12

quantify_5prime

429.542000000001s (~7.16 minutes)

1916

60

quantify_5prime

947.425999999992s (~15.79 minutes)

4174

120

quantify_5prime

1570.59999999999s (~26.18 minutes)

6017

180

quantify_5prime

2433.204s (~40.55 minutes)

7977

240

quantify_5prime

Mapping and quantification summary

prw_map <- bind_rows(prw_bt, prw_sr, prw_counts)

Running time and memory footprint depending on the number of reads:

p1 <- prw_map %>%
  ggplot() +
  geom_line(aes(x=nb_reads, y=time, color=features),
            show.legend = FALSE) +
  geom_point(aes(x=nb_reads, y=time, shape=features, color=features),
            show.legend = FALSE) +
  theme_light() +
  xlab("Number of reads (in millions)") +
  ylab("Duration (in seconds)") +
  ggtitle("Running times for mapping and quantification")

p2 <- prw_map %>%
  ggplot() +
  geom_line(aes(x=nb_reads, y=peak_ram, color=features)) +
  geom_point(aes(x=nb_reads, y=peak_ram, shape=features, color=features)) +
  theme_light() +
  xlab("Number of reads (in millions)") +
  ylab("Memory used (in MB)") +
  ggtitle("Memory used during mapping and quantification")  +
  theme(legend.position="inside", legend.position.inside = c(0.65, 0.2))
ggarrange(p1, p2) 

Note

In the above graphs, we can observe that the running times scale linearly with the number of sequencing reads to process.

Regarding the memory footprint, both aligners require a small amount of memory. The quantification to obtain the count table with the quantify_5prime function from rnaends requires an amount of memory that scales linearly with the number of reads as it has to process all reads to compute the 5’ end occurences.

TSS identification: TSS

Load quantifications by re-calling the last quantify_5prime with force=FALSE which will return the previous results.

i <- 5
bam_files <- list.files(
  paste0(
    list.files(  
      dirout[i],
      pattern = "_valid.fastq.gz$", 
      full.names = TRUE
      ),
    "_MW2.fasta_mm1_bowtie"
  ),
  pattern=".bam$",
  full.names = TRUE
)
count_table <- quantify_from_bam_files(
      bam_files = bam_files,
      sample_ids = samples,
      fun = quantify_5prime,
      force = FALSE
)
#> Quantification 5' output file exists, set force=T to overwrite: quantif.20M/prepro.PPP-BG.20M_AA_valid.fastq.gz_MW2.fasta_mm1_bowtie/prepro.PPP-BG.20M_AA_valid.fastq.gz.bam.csv
#> Quantification 5' output file exists, set force=T to overwrite: quantif.20M/prepro.PPP-BG.20M_AC_valid.fastq.gz_MW2.fasta_mm1_bowtie/prepro.PPP-BG.20M_AC_valid.fastq.gz.bam.csv
#> Quantification 5' output file exists, set force=T to overwrite: quantif.20M/prepro.PPP-BG.20M_AT_valid.fastq.gz_MW2.fasta_mm1_bowtie/prepro.PPP-BG.20M_AT_valid.fastq.gz.bam.csv
#> Quantification 5' output file exists, set force=T to overwrite: quantif.20M/prepro.PPP-BG.20M_CA_valid.fastq.gz_MW2.fasta_mm1_bowtie/prepro.PPP-BG.20M_CA_valid.fastq.gz.bam.csv
#> Quantification 5' output file exists, set force=T to overwrite: quantif.20M/prepro.PPP-BG.20M_CC_valid.fastq.gz_MW2.fasta_mm1_bowtie/prepro.PPP-BG.20M_CC_valid.fastq.gz.bam.csv
#> Quantification 5' output file exists, set force=T to overwrite: quantif.20M/prepro.PPP-BG.20M_CT_valid.fastq.gz_MW2.fasta_mm1_bowtie/prepro.PPP-BG.20M_CT_valid.fastq.gz.bam.csv

Design table

design_tb <- tibble(
  sample_id = samples,
  count_of = c("total", "total", "total", "fraction", "fraction", "fraction"),
  replicate = c(1,2,3, 1,2,3)
)
design_tb %>%
  knitr::kable()

sample_id

count_of

replicate

PPP.r1

total

1

PPP.r2

total

2

PPP.r3

total

3

BG.r1

fraction

1

BG.r2

fraction

2

BG.r3

fraction

3

Measure time and maximum RAM in use for the TSS function:

prw_tss <- sapply(1:length(dirout), function(i) {
  bam_files <- list.files(
    paste0(
      list.files(  
        dirout[i],
        pattern = "_valid.fastq.gz$", 
        full.names = TRUE
        ),
      "_MW2.fasta_mm1_bowtie"
    ),
    pattern=".bam$",
    full.names = TRUE
  )
  peakRAM(
    TSS(
      quantify_from_bam_files(
        bam_files = bam_files,
        sample_ids = samples,
        fun = quantify_5prime,
        force = FALSE
      ),
      design_tb))
  }
)
#> Quantification 5' output file exists, set force=T to overwrite: quantif.1M/prepro.PPP-BG.1M_AA_valid.fastq.gz_MW2.fasta_mm1_bowtie/prepro.PPP-BG.1M_AA_valid.fastq.gz.bam.csv
#> Quantification 5' output file exists, set force=T to overwrite: quantif.1M/prepro.PPP-BG.1M_AC_valid.fastq.gz_MW2.fasta_mm1_bowtie/prepro.PPP-BG.1M_AC_valid.fastq.gz.bam.csv
#> Quantification 5' output file exists, set force=T to overwrite: quantif.1M/prepro.PPP-BG.1M_AT_valid.fastq.gz_MW2.fasta_mm1_bowtie/prepro.PPP-BG.1M_AT_valid.fastq.gz.bam.csv
#> Quantification 5' output file exists, set force=T to overwrite: quantif.1M/prepro.PPP-BG.1M_CA_valid.fastq.gz_MW2.fasta_mm1_bowtie/prepro.PPP-BG.1M_CA_valid.fastq.gz.bam.csv
#> Quantification 5' output file exists, set force=T to overwrite: quantif.1M/prepro.PPP-BG.1M_CC_valid.fastq.gz_MW2.fasta_mm1_bowtie/prepro.PPP-BG.1M_CC_valid.fastq.gz.bam.csv
#> Quantification 5' output file exists, set force=T to overwrite: quantif.1M/prepro.PPP-BG.1M_CT_valid.fastq.gz_MW2.fasta_mm1_bowtie/prepro.PPP-BG.1M_CT_valid.fastq.gz.bam.csv
#> Quantification 5' output file exists, set force=T to overwrite: quantif.5M/prepro.PPP-BG.5M_AA_valid.fastq.gz_MW2.fasta_mm1_bowtie/prepro.PPP-BG.5M_AA_valid.fastq.gz.bam.csv
#> Quantification 5' output file exists, set force=T to overwrite: quantif.5M/prepro.PPP-BG.5M_AC_valid.fastq.gz_MW2.fasta_mm1_bowtie/prepro.PPP-BG.5M_AC_valid.fastq.gz.bam.csv
#> Quantification 5' output file exists, set force=T to overwrite: quantif.5M/prepro.PPP-BG.5M_AT_valid.fastq.gz_MW2.fasta_mm1_bowtie/prepro.PPP-BG.5M_AT_valid.fastq.gz.bam.csv
#> Quantification 5' output file exists, set force=T to overwrite: quantif.5M/prepro.PPP-BG.5M_CA_valid.fastq.gz_MW2.fasta_mm1_bowtie/prepro.PPP-BG.5M_CA_valid.fastq.gz.bam.csv
#> Quantification 5' output file exists, set force=T to overwrite: quantif.5M/prepro.PPP-BG.5M_CC_valid.fastq.gz_MW2.fasta_mm1_bowtie/prepro.PPP-BG.5M_CC_valid.fastq.gz.bam.csv
#> Quantification 5' output file exists, set force=T to overwrite: quantif.5M/prepro.PPP-BG.5M_CT_valid.fastq.gz_MW2.fasta_mm1_bowtie/prepro.PPP-BG.5M_CT_valid.fastq.gz.bam.csv
#> Quantification 5' output file exists, set force=T to overwrite: quantif.10M/prepro.PPP-BG.10M_AA_valid.fastq.gz_MW2.fasta_mm1_bowtie/prepro.PPP-BG.10M_AA_valid.fastq.gz.bam.csv
#> Quantification 5' output file exists, set force=T to overwrite: quantif.10M/prepro.PPP-BG.10M_AC_valid.fastq.gz_MW2.fasta_mm1_bowtie/prepro.PPP-BG.10M_AC_valid.fastq.gz.bam.csv
#> Quantification 5' output file exists, set force=T to overwrite: quantif.10M/prepro.PPP-BG.10M_AT_valid.fastq.gz_MW2.fasta_mm1_bowtie/prepro.PPP-BG.10M_AT_valid.fastq.gz.bam.csv
#> Quantification 5' output file exists, set force=T to overwrite: quantif.10M/prepro.PPP-BG.10M_CA_valid.fastq.gz_MW2.fasta_mm1_bowtie/prepro.PPP-BG.10M_CA_valid.fastq.gz.bam.csv
#> Quantification 5' output file exists, set force=T to overwrite: quantif.10M/prepro.PPP-BG.10M_CC_valid.fastq.gz_MW2.fasta_mm1_bowtie/prepro.PPP-BG.10M_CC_valid.fastq.gz.bam.csv
#> Quantification 5' output file exists, set force=T to overwrite: quantif.10M/prepro.PPP-BG.10M_CT_valid.fastq.gz_MW2.fasta_mm1_bowtie/prepro.PPP-BG.10M_CT_valid.fastq.gz.bam.csv
#> Quantification 5' output file exists, set force=T to overwrite: quantif.15M/prepro.PPP-BG.15M_AA_valid.fastq.gz_MW2.fasta_mm1_bowtie/prepro.PPP-BG.15M_AA_valid.fastq.gz.bam.csv
#> Quantification 5' output file exists, set force=T to overwrite: quantif.15M/prepro.PPP-BG.15M_AC_valid.fastq.gz_MW2.fasta_mm1_bowtie/prepro.PPP-BG.15M_AC_valid.fastq.gz.bam.csv
#> Quantification 5' output file exists, set force=T to overwrite: quantif.15M/prepro.PPP-BG.15M_AT_valid.fastq.gz_MW2.fasta_mm1_bowtie/prepro.PPP-BG.15M_AT_valid.fastq.gz.bam.csv
#> Quantification 5' output file exists, set force=T to overwrite: quantif.15M/prepro.PPP-BG.15M_CA_valid.fastq.gz_MW2.fasta_mm1_bowtie/prepro.PPP-BG.15M_CA_valid.fastq.gz.bam.csv
#> Quantification 5' output file exists, set force=T to overwrite: quantif.15M/prepro.PPP-BG.15M_CC_valid.fastq.gz_MW2.fasta_mm1_bowtie/prepro.PPP-BG.15M_CC_valid.fastq.gz.bam.csv
#> Quantification 5' output file exists, set force=T to overwrite: quantif.15M/prepro.PPP-BG.15M_CT_valid.fastq.gz_MW2.fasta_mm1_bowtie/prepro.PPP-BG.15M_CT_valid.fastq.gz.bam.csv
#> Quantification 5' output file exists, set force=T to overwrite: quantif.20M/prepro.PPP-BG.20M_AA_valid.fastq.gz_MW2.fasta_mm1_bowtie/prepro.PPP-BG.20M_AA_valid.fastq.gz.bam.csv
#> Quantification 5' output file exists, set force=T to overwrite: quantif.20M/prepro.PPP-BG.20M_AC_valid.fastq.gz_MW2.fasta_mm1_bowtie/prepro.PPP-BG.20M_AC_valid.fastq.gz.bam.csv
#> Quantification 5' output file exists, set force=T to overwrite: quantif.20M/prepro.PPP-BG.20M_AT_valid.fastq.gz_MW2.fasta_mm1_bowtie/prepro.PPP-BG.20M_AT_valid.fastq.gz.bam.csv
#> Quantification 5' output file exists, set force=T to overwrite: quantif.20M/prepro.PPP-BG.20M_CA_valid.fastq.gz_MW2.fasta_mm1_bowtie/prepro.PPP-BG.20M_CA_valid.fastq.gz.bam.csv
#> Quantification 5' output file exists, set force=T to overwrite: quantif.20M/prepro.PPP-BG.20M_CC_valid.fastq.gz_MW2.fasta_mm1_bowtie/prepro.PPP-BG.20M_CC_valid.fastq.gz.bam.csv
#> Quantification 5' output file exists, set force=T to overwrite: quantif.20M/prepro.PPP-BG.20M_CT_valid.fastq.gz_MW2.fasta_mm1_bowtie/prepro.PPP-BG.20M_CT_valid.fastq.gz.bam.csv
prw_tss <- prw_tss %>%
  t %>%
  as_tibble() %>%
  select(time=Elapsed_Time_sec, 
         peak_ram=Peak_RAM_Used_MiB) %>%
  mutate(time=duration(as.numeric(time), units="seconds"), 
         peak_ram = as.numeric(peak_ram),
         nb_reads = c(12, 60, 120, 180, 240),
         analysis = "TSS")
prw_tss %>%
  knitr::kable()

time

peak_ram

nb_reads

features

4.18499999999767s

2206

12

TSS

25.1370000000024s

1263

60

TSS

52.4130000000005s

924

120

TSS

75.1419999999925s (~1.25 minutes)

924

180

TSS

93.3399999999965s (~1.56 minutes)

924

240

TSS

Summary

p1 <- prw_tss %>%
  ggplot() +
  geom_line(aes(x=nb_reads, y=time, color=features),
            show.legend = FALSE) +
  geom_point(aes(x=nb_reads, y=time, shape=features, color=features),
            show.legend = FALSE) +
  theme_light() +
  xlab("Number of reads (in millions)") +
  ylab("Duration (in seconds)") +
  ggtitle("Running times for TSS identification")

p2 <- prw_tss %>%
  ggplot() +
  geom_line(aes(x=nb_reads, y=peak_ram, color=features),
            show.legend = FALSE) +
  geom_point(aes(x=nb_reads, y=peak_ram, shape=features, color=features),
            show.legend = FALSE) +
  theme_light() +
  xlab("Number of reads (in millions)") +
  ylab("Memory used (in MB)") +
  ggtitle("Memory used during TSS identification")
ggarrange(p1, p2) 

Note

In the above graphs, we can observe that the running times scale linearly with the number of reads processed. Regarding the memory footprint, once the count table is computed in the previous analysis step, we can observe that the tibble representation used its storage is around 1-2GB.

mem_used()
#> 1.15 GB