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fastpのバッチ処理スクリプトを使う

2025/09/19 追記

 

fastpのversion 1.0がリリースされ、フォルダ内のfastqをバッチ処理する便利なスクリプトも提供されました。このスクリプトの使い方を確認しておきます。

 

インストール

最新のfastpにパスが通っている必要がある。fastpのバージョンが1未満だと動作しない。

Github

#bioconda (link) v.1系指定、最新バージョンはリンク先確認
mamba install -c bioconda -y fastp==1.0.1
# download the latest build
wget http://opengene.org/fastp/fastp
chmod a+x ./fastp

#parallel.pyはcondaではインストールされない。レポジトリから取得する。
git clone https://github.com/OpenGene/fastp.git
#パスの通ったディレクトリに移動させる
chmod +x fastp/parallel.py
mv fastp/parallel.py /usr/local/bin/ #(あるいは~/binなど)

> python parallel.py -h

Usage: A python script to use fastp to preprocess all FASTQ files within a folder

 

Options:

  --version             show program's version number and exit

  -h, --help            show this help message and exit

  -i INPUT_DIR, --input_dir=INPUT_DIR

                        the folder contains the FASTQ files to be

                        preprocessed, by default is current dir (.)

  -o OUT_DIR, --out_dir=OUT_DIR

                        the folder to store the clean FASTQ. If not specified,

                        then there will be no output files.

  -r REPORT_DIR, --report_dir=REPORT_DIR

                        the folder to store QC reports. If not specified, use

                        out_dir if out_dir is specified, otherwise use

                        input_dir.

  -c COMMAND, --command=COMMAND

                        the path to fastp command, if not specified, then it

                        will use 'fastp' in PATH

  -a ARGS, --args=ARGS  the arguments that will be passed to fastp. Enclose in

                        quotation marks. Like --args='-f 3 -t 3'

  -p PARALLEL, --parallel=PARALLEL

                        the number of fastp processes can be run in parallel,

                        if not specified, then it will be CPU_Core/4

  -1 READ1_FLAG, --read1_flag=READ1_FLAG

                        specify the name flag of read1, default is R1, which

                        means a file with name *R1* is read1 file

  -2 READ2_FLAG, --read2_flag=READ2_FLAG

                        specify the name flag of read2, default is R2, which

                        means a file with name *R2* is read2 file

 

 

実行方法

fastq(fastq.gz)を含むディレクトリ、出力ディレクトリ、HTMLとJSON形式レポートの保存ディレクトリをそれぞれ指定する。fastpのネイティブオプションのコマンドは-a ' 'で指定する。例えば-f3 -t2は各fastqの先頭3bpと末尾2bpをそれぞれ強制トリミングする。

python parallel.py -i input_fastq_dir -o fq_outdir -r report_outdir -a '-f 3 -t 2'
  • -i     the folder contains the FASTQ files to be preprocessed, by default is current dir (.)
  • -o    the folder to store the clean FASTQ. If not specified, then there will be no output files.
  • -r     the folder to store QC reports. If not specified, use out_dir if out_dir is specified, otherwise use input_dir. 
  • -a     the arguments that will be passed to fastp. Enclose in quotation marks. Like --args='-f 3 -t 3'
  •  -p    the number of fastp processes can be run in parallel, if not specified, then it will be CPU_Core/4
  •  -1     specify the name flag of read1, default is R1, which means a file with name *R1* is read1 file
  • -2     specify the name flag of read2, default is R2, which means a file with name *R2* is read2 file
                            

結果は指定したフォルダ内に保存される。ペアエンドのリードはデフォルトではR1R2が認識される。変更するには-1-2オプションを使う。

parallel.py \
-i ./ -o fastp_processed -r report -p 6 \
-1 "_R1" -2 "_R2" \
-a "-q 20 -u 30 -n 10 -l 20 --correction -w 3"
  • -c, --correction     enable base correction in overlapped regions (only for PE data), default is disabled

  •  

     -u, --unqualified_percent_limit      how many percents of bases are allowed to be unqualified (0~100). Default 40 means 40% (int [=40])     

 

その他

  • ペアエンドの片方しか認識されなかったりルールとファイル名が異なる場合、スキップされるわけではなく2つのシングルエンドとして処理される(同期が取れなくなっている可能性が高いので注意)。
  • この挙動から、シングルエンドとペアエンドが混在していても全てのfastqをバッチ処理できると考えられる。

引用

fastp 1.0: An ultra-fast all-round tool for FASTQ data quality control and preprocessing

Shifu Chen

iMeta, First published: 09 September 2025. 

 

関連

 

v1.0のoptipn

$ fastp

fastp: an ultra-fast all-in-one FASTQ preprocessor

version 1.0.1

usage: fastp [options] ... 

options:

  -i, --in1                            read1 input file name (string [=])

  -o, --out1                           read1 output file name (string [=])

  -I, --in2                            read2 input file name (string [=])

  -O, --out2                           read2 output file name (string [=])

      --unpaired1                      for PE input, if read1 passed QC but read2 not, it will be written to unpaired1. Default is to discard it. (string [=])

      --unpaired2                      for PE input, if read2 passed QC but read1 not, it will be written to unpaired2. If --unpaired2 is same as --unpaired1 (default mode), both unpaired reads will be written to this same file. (string [=])

      --overlapped_out                 for each read pair, output the overlapped region if it has no any mismatched base. (string [=])

      --failed_out                     specify the file to store reads that cannot pass the filters. (string [=])

  -m, --merge                          for paired-end input, merge each pair of reads into a single read if they are overlapped. The merged reads will be written to the file given by --merged_out, the unmerged reads will be written to the files specified by --out1 and --out2. The merging mode is disabled by default.

      --merged_out                     in the merging mode, specify the file name to store merged output, or specify --stdout to stream the merged output (string [=])

      --include_unmerged               in the merging mode, write the unmerged or unpaired reads to the file specified by --merge. Disabled by default.

  -6, --phred64                        indicate the input is using phred64 scoring (it'll be converted to phred33, so the output will still be phred33)

  -z, --compression                    compression level for gzip output (1 ~ 9). 1 is fastest, 9 is smallest, default is 4. (int [=4])

      --stdin                          input from STDIN. If the STDIN is interleaved paired-end FASTQ, please also add --interleaved_in.

      --stdout                         stream passing-filters reads to STDOUT. This option will result in interleaved FASTQ output for paired-end output. Disabled by default.

      --interleaved_in                 indicate that <in1> is an interleaved FASTQ which contains both read1 and read2. Disabled by default.

      --reads_to_process               specify how many reads/pairs to be processed. Default 0 means process all reads. (int [=0])

      --dont_overwrite                 don't overwrite existing files. Overwritting is allowed by default.

      --fix_mgi_id                     the MGI FASTQ ID format is not compatible with many BAM operation tools, enable this option to fix it.

  -V, --verbose                        output verbose log information (i.e. when every 1M reads are processed).

  -A, --disable_adapter_trimming       adapter trimming is enabled by default. If this option is specified, adapter trimming is disabled

  -a, --adapter_sequence               the adapter for read1. For SE data, if not specified, the adapter will be auto-detected. For PE data, this is used if R1/R2 are found not overlapped. (string [=auto])

      --adapter_sequence_r2            the adapter for read2 (PE data only). This is used if R1/R2 are found not overlapped. If not specified, it will be the same as <adapter_sequence> (string [=auto])

      --adapter_fasta                  specify a FASTA file to trim both read1 and read2 (if PE) by all the sequences in this FASTA file (string [=])

  -2, --detect_adapter_for_pe          enable adapter detection for PE data to get ultra-clean data. It takes more time to find just a little bit more adapters.

      --allow_gap_overlap_trimming     allow up to one gap when trim adapters by overlap analysis for PE data. By default no gap is allowed.

  -f, --trim_front1                    trimming how many bases in front for read1, default is 0 (int [=0])

  -t, --trim_tail1                     trimming how many bases in tail for read1, default is 0 (int [=0])

  -b, --max_len1                       if read1 is longer than max_len1, then trim read1 at its tail to make it as long as max_len1. Default 0 means no limitation (int [=0])

  -F, --trim_front2                    trimming how many bases in front for read2. If it's not specified, it will follow read1's settings (int [=0])

  -T, --trim_tail2                     trimming how many bases in tail for read2. If it's not specified, it will follow read1's settings (int [=0])

  -B, --max_len2                       if read2 is longer than max_len2, then trim read2 at its tail to make it as long as max_len2. Default 0 means no limitation. If it's not specified, it will follow read1's settings (int [=0])

  -D, --dedup                          enable deduplication to drop the duplicated reads/pairs

      --dup_calc_accuracy              accuracy level to calculate duplication (1~6), higher level uses more memory (1G, 2G, 4G, 8G, 16G, 24G). Default 1 for no-dedup mode, and 3 for dedup mode. (int [=0])

      --dont_eval_duplication          don't evaluate duplication rate to save time and use less memory.

  -g, --trim_poly_g                    force polyG tail trimming, by default trimming is automatically enabled for Illumina NextSeq/NovaSeq data

      --poly_g_min_len                 the minimum length to detect polyG in the read tail. 10 by default. (int [=10])

  -G, --disable_trim_poly_g            disable polyG tail trimming, by default trimming is automatically enabled for Illumina NextSeq/NovaSeq data

  -x, --trim_poly_x                    enable polyX trimming in 3' ends.

      --poly_x_min_len                 the minimum length to detect polyX in the read tail. 10 by default. (int [=10])

  -5, --cut_front                      move a sliding window from front (5') to tail, drop the bases in the window if its mean quality < threshold, stop otherwise.

  -3, --cut_tail                       move a sliding window from tail (3') to front, drop the bases in the window if its mean quality < threshold, stop otherwise.

  -r, --cut_right                      move a sliding window from front to tail, if meet one window with mean quality < threshold, drop the bases in the window and the right part, and then stop.

  -W, --cut_window_size                the window size option shared by cut_front, cut_tail or cut_sliding. Range: 1~1000, default: 4 (int [=4])

  -M, --cut_mean_quality               the mean quality requirement option shared by cut_front, cut_tail or cut_sliding. Range: 1~36 default: 20 (Q20) (int [=20])

      --cut_front_window_size          the window size option of cut_front, default to cut_window_size if not specified (int [=4])

      --cut_front_mean_quality         the mean quality requirement option for cut_front, default to cut_mean_quality if not specified (int [=20])

      --cut_tail_window_size           the window size option of cut_tail, default to cut_window_size if not specified (int [=4])

      --cut_tail_mean_quality          the mean quality requirement option for cut_tail, default to cut_mean_quality if not specified (int [=20])

      --cut_right_window_size          the window size option of cut_right, default to cut_window_size if not specified (int [=4])

      --cut_right_mean_quality         the mean quality requirement option for cut_right, default to cut_mean_quality if not specified (int [=20])

  -Q, --disable_quality_filtering      quality filtering is enabled by default. If this option is specified, quality filtering is disabled

  -q, --qualified_quality_phred        the quality value that a base is qualified. Default 15 means phred quality >=Q15 is qualified. (int [=15])

  -u, --unqualified_percent_limit      how many percents of bases are allowed to be unqualified (0~100). Default 40 means 40% (int [=40])

  -n, --n_base_limit                   if one read's number of N base is >n_base_limit, then this read/pair is discarded. Default is 5 (int [=5])

  -e, --average_qual                   if one read's average quality score <avg_qual, then this read/pair is discarded. Default 0 means no requirement (int [=0])

  -L, --disable_length_filtering       length filtering is enabled by default. If this option is specified, length filtering is disabled

  -l, --length_required                reads shorter than length_required will be discarded, default is 15. (int [=15])

      --length_limit                   reads longer than length_limit will be discarded, default 0 means no limitation. (int [=0])

  -y, --low_complexity_filter          enable low complexity filter. The complexity is defined as the percentage of base that is different from its next base (base[i] != base[i+1]).

  -Y, --complexity_threshold           the threshold for low complexity filter (0~100). Default is 30, which means 30% complexity is required. (int [=30])

      --filter_by_index1               specify a file contains a list of barcodes of index1 to be filtered out, one barcode per line (string [=])

      --filter_by_index2               specify a file contains a list of barcodes of index2 to be filtered out, one barcode per line (string [=])

      --filter_by_index_threshold      the allowed difference of index barcode for index filtering, default 0 means completely identical. (int [=0])

  -c, --correction                     enable base correction in overlapped regions (only for PE data), default is disabled

      --overlap_len_require            the minimum length to detect overlapped region of PE reads. This will affect overlap analysis based PE merge, adapter trimming and correction. 30 by default. (int [=30])

      --overlap_diff_limit             the maximum number of mismatched bases to detect overlapped region of PE reads. This will affect overlap analysis based PE merge, adapter trimming and correction. 5 by default. (int [=5])

      --overlap_diff_percent_limit     the maximum percentage of mismatched bases to detect overlapped region of PE reads. This will affect overlap analysis based PE merge, adapter trimming and correction. Default 20 means 20%. (int [=20])

  -U, --umi                            enable unique molecular identifier (UMI) preprocessing

      --umi_loc                        specify the location of UMI, can be (index1/index2/read1/read2/per_index/per_read, default is none (string [=])

      --umi_len                        if the UMI is in read1/read2, its length should be provided (int [=0])

      --umi_prefix                     if specified, an underline will be used to connect prefix and UMI (i.e. prefix=UMI, UMI=AATTCG, final=UMI_AATTCG). No prefix by default (string [=])

      --umi_skip                       if the UMI is in read1/read2, fastp can skip several bases following UMI, default is 0 (int [=0])

      --umi_delim                      delimiter to use between the read name and the UMI, default is : (string [=:])

  -p, --overrepresentation_analysis    enable overrepresented sequence analysis.

  -P, --overrepresentation_sampling    one in (--overrepresentation_sampling) reads will be computed for overrepresentation analysis (1~10000), smaller is slower, default is 20. (int [=20])

  -j, --json                           the json format report file name (string [=fastp.json])

  -h, --html                           the html format report file name (string [=fastp.html])

  -R, --report_title                   should be quoted with ' or ", default is "fastp report" (string [=fastp report])

  -w, --thread                         worker thread number, default is 3 (int [=3])

  -s, --split                          split output by limiting total split file number with this option (2~999), a sequential number prefix will be added to output name ( 0001.out.fq, 0002.out.fq...), disabled by default (int [=0])

  -S, --split_by_lines                 split output by limiting lines of each file with this option(>=1000), a sequential number prefix will be added to output name ( 0001.out.fq, 0002.out.fq...), disabled by default (long [=0])

  -d, --split_prefix_digits            the digits for the sequential number padding (1~10), default is 4, so the filename will be padded as 0001.xxx, 0 to disable padding (int [=4])

      --cut_by_quality5                DEPRECATED, use --cut_front instead.

      --cut_by_quality3                DEPRECATED, use --cut_tail instead.

      --cut_by_quality_aggressive      DEPRECATED, use --cut_right instead.

      --discard_unmerged               DEPRECATED, no effect now, see the introduction for merging.

  -?, --help                           print this message

 

Citation:

Shifu Chen. 2023. Ultrafast one-pass FASTQ data preprocessing, quality control, and deduplication using fastp. iMeta 2: e107