.\" DO NOT MODIFY THIS FILE! It was generated by help2man 1.48.2. .TH BOWTIE2 "1" "March 2021" "bowtie2 2.4.2" "User Commands" .SH NAME bowtie2 \- wrapper for bowtie2-align-* .SH DESCRIPTION Bowtie 2 version 2.4.2 by Ben Langmead (langmea@cs.jhu.edu, www.cs.jhu.edu/~langmea) .SH USAGE bowtie2 [options]* \fB\-x\fR {\-1 \fB\-2\fR | \fB\-U\fR | \fB\-\-interleaved\fR | \fB\-b\fR } [\-S ] .TP Index filename prefix (minus trailing .X.bt2). NOTE: Bowtie 1 and Bowtie 2 indexes are not compatible. .TP Files with #1 mates, paired with files in . Could be gzip'ed (extension: .gz) or bzip2'ed (extension: .bz2). .TP Files with #2 mates, paired with files in . Could be gzip'ed (extension: .gz) or bzip2'ed (extension: .bz2). .TP Files with unpaired reads. Could be gzip'ed (extension: .gz) or bzip2'ed (extension: .bz2). .TP Files with interleaved paired\-end FASTQ/FASTA reads Could be gzip'ed (extension: .gz) or bzip2'ed (extension: .bz2). .TP Files are unaligned BAM sorted by read name. .TP File for SAM output (default: stdout) .IP , , can be comma\-separated lists (no whitespace) and can be specified many times. E.g. '\-U file1.fq,file2.fq \fB\-U\fR file3.fq'. .PP .SH OPTIONS (defaults in parentheses) .SS Input: .TP \fB\-q\fR query input files are FASTQ .fq/.fastq (default) .TP \fB\-\-tab5\fR query input files are TAB5 .tab5 .TP \fB\-\-tab6\fR query input files are TAB6 .tab6 .TP \fB\-\-qseq\fR query input files are in Illumina's qseq format .TP \fB\-f\fR query input files are (multi\-)FASTA .fa/.mfa .TP \fB\-r\fR query input files are raw one\-sequence\-per\-line .TP \fB\-F\fR k:,i: query input files are continuous FASTA where reads are substrings (k\-mers) extracted from a FASTA file and aligned at offsets 1, 1+i, 1+2i ... end of reference .TP \fB\-c\fR , , are sequences themselves, not files .TP \fB\-s\fR/\-\-skip skip the first reads/pairs in the input (none) .TP \fB\-u\fR/\-\-upto stop after first reads/pairs (no limit) .TP \fB\-5\fR/\-\-trim5 trim bases from 5'/left end of reads (0) .TP \fB\-3\fR/\-\-trim3 trim bases from 3'/right end of reads (0) .TP \fB\-\-trim\-to\fR [3:|5:] trim reads exceeding bases from either 3' or 5' end If the read end is not specified then it defaults to 3 (0) .TP \fB\-\-phred33\fR qualities are Phred+33 (default) .TP \fB\-\-phred64\fR qualities are Phred+64 .TP \fB\-\-int\-quals\fR qualities encoded as space\-delimited integers .SS Presets: Same as: .IP For \fB\-\-end\-to\-end\fR: .HP \fB\-\-very\-fast\fR \fB\-D\fR 5 \fB\-R\fR 1 \fB\-N\fR 0 \fB\-L\fR 22 \fB\-i\fR S,0,2.50 .HP \fB\-\-fast\fR \fB\-D\fR 10 \fB\-R\fR 2 \fB\-N\fR 0 \fB\-L\fR 22 \fB\-i\fR S,0,2.50 .HP \fB\-\-sensitive\fR \fB\-D\fR 15 \fB\-R\fR 2 \fB\-N\fR 0 \fB\-L\fR 22 \fB\-i\fR S,1,1.15 (default) .HP \fB\-\-very\-sensitive\fR \fB\-D\fR 20 \fB\-R\fR 3 \fB\-N\fR 0 \fB\-L\fR 20 \fB\-i\fR S,1,0.50 .IP For \fB\-\-local\fR: .HP \fB\-\-very\-fast\-local\fR \fB\-D\fR 5 \fB\-R\fR 1 \fB\-N\fR 0 \fB\-L\fR 25 \fB\-i\fR S,1,2.00 .HP \fB\-\-fast\-local\fR \fB\-D\fR 10 \fB\-R\fR 2 \fB\-N\fR 0 \fB\-L\fR 22 \fB\-i\fR S,1,1.75 .HP \fB\-\-sensitive\-local\fR \fB\-D\fR 15 \fB\-R\fR 2 \fB\-N\fR 0 \fB\-L\fR 20 \fB\-i\fR S,1,0.75 (default) .HP \fB\-\-very\-sensitive\-local\fR \fB\-D\fR 20 \fB\-R\fR 3 \fB\-N\fR 0 \fB\-L\fR 20 \fB\-i\fR S,1,0.50 .SS Alignment: .TP \fB\-N\fR max # mismatches in seed alignment; can be 0 or 1 (0) .TP \fB\-L\fR length of seed substrings; must be >3, <32 (22) .TP \fB\-i\fR interval between seed substrings w/r/t read len (S,1,1.15) .TP \fB\-\-n\-ceil\fR func for max # non\-A/C/G/Ts permitted in aln (L,0,0.15) .TP \fB\-\-dpad\fR include extra ref chars on sides of DP table (15) .TP \fB\-\-gbar\fR disallow gaps within nucs of read extremes (4) .TP \fB\-\-ignore\-quals\fR treat all quality values as 30 on Phred scale (off) .TP \fB\-\-nofw\fR do not align forward (original) version of read (off) .TP \fB\-\-norc\fR do not align reverse\-complement version of read (off) .TP \fB\-\-no\-1mm\-upfront\fR do not allow 1 mismatch alignments before attempting to scan for the optimal seeded alignments .TP \fB\-\-end\-to\-end\fR entire read must align; no clipping (on) .IP OR .TP \fB\-\-local\fR local alignment; ends might be soft clipped (off) .SS Scoring: .TP \fB\-\-ma\fR match bonus (0 for \fB\-\-end\-to\-end\fR, 2 for \fB\-\-local\fR) .TP \fB\-\-mp\fR max penalty for mismatch; lower qual = lower penalty (6) .TP \fB\-\-np\fR penalty for non\-A/C/G/Ts in read/ref (1) .TP \fB\-\-rdg\fR , read gap open, extend penalties (5,3) .TP \fB\-\-rfg\fR , reference gap open, extend penalties (5,3) .TP \fB\-\-score\-min\fR min acceptable alignment score w/r/t read length (G,20,8 for local, L,\-0.6,\-0.6 for end\-to\-end) .SS Reporting: .TP (default) look for multiple alignments, report best, with MAPQ .IP OR .TP \fB\-k\fR report up to alns per read; MAPQ not meaningful .IP OR .TP \fB\-a\fR/\-\-all report all alignments; very slow, MAPQ not meaningful .SS Effort: .TP \fB\-D\fR give up extending after failed extends in a row (15) .TP \fB\-R\fR for reads w/ repetitive seeds, try sets of seeds (2) .IP Paired\-end: .TP \fB\-I\fR/\-\-minins minimum fragment length (0) .TP \fB\-X\fR/\-\-maxins maximum fragment length (500) .HP \fB\-\-fr\fR/\-\-rf/\-\-ff \fB\-1\fR, \fB\-2\fR mates align fw/rev, rev/fw, fw/fw (\fB\-\-fr\fR) .TP \fB\-\-no\-mixed\fR suppress unpaired alignments for paired reads .TP \fB\-\-no\-discordant\fR suppress discordant alignments for paired reads .TP \fB\-\-dovetail\fR concordant when mates extend past each other .TP \fB\-\-no\-contain\fR not concordant when one mate alignment contains other .TP \fB\-\-no\-overlap\fR not concordant when mates overlap at all .IP BAM: .TP \fB\-\-align\-paired\-reads\fR Bowtie2 will, by default, attempt to align unpaired BAM reads. Use this option to align paired\-end reads instead. .TP \fB\-\-preserve\-tags\fR Preserve tags from the original BAM record by appending them to the end of the corresponding SAM output. .SS Output: .TP \fB\-t\fR/\-\-time print wall\-clock time taken by search phases .TP \fB\-\-un\fR write unpaired reads that didn't align to .TP \fB\-\-al\fR write unpaired reads that aligned at least once to .TP \fB\-\-un\-conc\fR write pairs that didn't align concordantly to .TP \fB\-\-al\-conc\fR write pairs that aligned concordantly at least once to .IP (Note: for \fB\-\-un\fR, \fB\-\-al\fR, \fB\-\-un\-conc\fR, or \fB\-\-al\-conc\fR, add '\-gz' to the option name, e.g. \fB\-\-un\-gz\fR , to gzip compress output, or add '\-bz2' to bzip2 compress output.) .TP \fB\-\-quiet\fR print nothing to stderr except serious errors .TP \fB\-\-met\-file\fR send metrics to file at (off) .TP \fB\-\-met\-stderr\fR send metrics to stderr (off) .TP \fB\-\-met\fR report internal counters & metrics every secs (1) .TP \fB\-\-no\-unal\fR suppress SAM records for unaligned reads .TP \fB\-\-no\-head\fR suppress header lines, i.e. lines starting with @ .TP \fB\-\-no\-sq\fR suppress @SQ header lines .TP \fB\-\-rg\-id\fR set read group id, reflected in @RG line and RG:Z: opt field .TP \fB\-\-rg\fR add ("lab:value") to @RG line of SAM header. Note: @RG line only printed when \fB\-\-rg\-id\fR is set. .TP \fB\-\-omit\-sec\-seq\fR put '*' in SEQ and QUAL fields for secondary alignments. .TP \fB\-\-sam\-no\-qname\-trunc\fR Suppress standard behavior of truncating readname at first whitespace at the expense of generating non\-standard SAM. .TP \fB\-\-xeq\fR Use '='/'X', instead of 'M,' to specify matches/mismatches in SAM record. .TP \fB\-\-soft\-clipped\-unmapped\-tlen\fR Exclude soft\-clipped bases when reporting TLEN .TP \fB\-\-sam\-append\-comment\fR Append FASTA/FASTQ comment to SAM record .SS Performance: .HP \fB\-p\fR/\-\-threads number of alignment threads to launch (1) .TP \fB\-\-reorder\fR force SAM output order to match order of input reads .TP \fB\-\-mm\fR use memory\-mapped I/O for index; many 'bowtie's can share .SS Other: .TP \fB\-\-qc\-filter\fR filter out reads that are bad according to QSEQ filter .TP \fB\-\-seed\fR seed for random number generator (0) .TP \fB\-\-non\-deterministic\fR seed rand. gen. arbitrarily instead of using read attributes .TP \fB\-\-version\fR print version information and quit .TP \fB\-h\fR/\-\-help print this usage message .PP