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howto:bioinformatics:bowtie

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Running Bowtie

Before you start

Before running your scripts remember to run1):

user@login01:~ $ module add bowtie/1.0.0

Job script

Then your job script called my_job.msub will look something like this:

my_job.msub
#/bin/sh
#MSUB -l procs=64
#MSUB -l feature=dell
#MSUB -l walltime=10:00:00
#MSUB -m be
#MSUB -V
#MSUB -o ${HOME}/scratch5/BOWTIE/stdout.txt
#MSUB -e ${HOME}/scratch5/BOWTIE/stderr.txt
#MSUB -d ${HOME}/scratch5/BOWTIE/
#MSUB -N MyBowtieJob
#MSUB -mb
 
NP=`cat ${PBS_NODEFILE} | wc -l`
 
EXE=bowtie
ARGS="sscrofa --shmem - p ${NP} -1 A_reads1,B_reads_1 -2 A_reads2,B_reads2 piggy_hits"
 
${EXE} ${ARGS}

Submit your job

Finally submit your job using:

user@login01:~ $ msub my_job.msub

Bowtie help

obtained by running:

user@login01:~ $ bowtie -h

Usage

  bowtie [options]* <ebwt> {-1 <m1> -2 <m2> | --12 <r> | <s>} [<hit>]

  <m1>    Comma-separated list of files containing upstream mates (or the
          sequences themselves, if -c is set) paired with mates in <m2>
  <m2>    Comma-separated list of files containing downstream mates (or the
          sequences themselves if -c is set) paired with mates in <m1>
  <r>     Comma-separated list of files containing Crossbow-style reads.  Can be
          a mixture of paired and unpaired.  Specify "-" for stdin.
  <s>     Comma-separated list of files containing unpaired reads, or the
          sequences themselves, if -c is set.  Specify "-" for stdin.
  <hit>   File to write hits to (default: stdout)

Options

Input

  -q                 query input files are FASTQ .fq/.fastq (default)
  -f                 query input files are (multi-)FASTA .fa/.mfa
  -r                 query input files are raw one-sequence-per-line
  -c                 query sequences given on cmd line (as <mates>, <singles>)
  -C                 reads and index are in colorspace
  -Q/--quals <file>  QV file(s) corresponding to CSFASTA inputs; use with -f -C
  --Q1/--Q2 <file>   same as -Q, but for mate files 1 and 2 respectively
  -s/--skip <int>    skip the first <int> reads/pairs in the input
  -u/--qupto <int>   stop after first <int> reads/pairs (excl. skipped reads)
  -5/--trim5 <int>   trim <int> bases from 5' (left) end of reads
  -3/--trim3 <int>   trim <int> bases from 3' (right) end of reads
  --phred33-quals    input quals are Phred+33 (default)
  --phred64-quals    input quals are Phred+64 (same as --solexa1.3-quals)
  --solexa-quals     input quals are from GA Pipeline ver. < 1.3
  --solexa1.3-quals  input quals are from GA Pipeline ver. >= 1.3
  --integer-quals    qualities are given as space-separated integers (not ASCII)

Alignment

  -v <int>           report end-to-end hits w/ <=v mismatches; ignore qualities
    or
  -n/--seedmms <int> max mismatches in seed (can be 0-3, default: -n 2)
  -e/--maqerr <int>  max sum of mismatch quals across alignment for -n (def: 70)
  -l/--seedlen <int> seed length for -n (default: 28)
  --nomaqround       disable Maq-like quality rounding for -n (nearest 10 <= 30)
  -I/--minins <int>  minimum insert size for paired-end alignment (default: 0)
  -X/--maxins <int>  maximum insert size for paired-end alignment (default: 250)
  --fr/--rf/--ff     -1, -2 mates align fw/rev, rev/fw, fw/fw (default: --fr)
  --nofw/--norc      do not align to forward/reverse-complement reference strand
  --maxbts <int>     max # backtracks for -n 2/3 (default: 125, 800 for --best)
  --pairtries <int>  max # attempts to find mate for anchor hit (default: 100)
  -y/--tryhard       try hard to find valid alignments, at the expense of speed
  --chunkmbs <int>   max megabytes of RAM for best-first search frames (def: 64)

Reporting

  -k <int>           report up to <int> good alignments per read (default: 1)
  -a/--all           report all alignments per read (much slower than low -k)
  -m <int>           suppress all alignments if > <int> exist (def: no limit)
  -M <int>           like -m, but reports 1 random hit (MAPQ=0); requires --best
  --best             hits guaranteed best stratum; ties broken by quality
  --strata           hits in sub-optimal strata aren't reported (requires --best)

Output

  -t/--time          print wall-clock time taken by search phases
  -B/--offbase <int> leftmost ref offset = <int> in bowtie output (default: 0)
  --quiet            print nothing but the alignments
  --refout           write alignments to files refXXXXX.map, 1 map per reference
  --refidx           refer to ref. seqs by 0-based index rather than name
  --al <fname>       write aligned reads/pairs to file(s) <fname>
  --un <fname>       write unaligned reads/pairs to file(s) <fname>
  --max <fname>      write reads/pairs over -m limit to file(s) <fname>
  --suppress <cols>  suppresses given columns (comma-delim'ed) in default output
  --fullref          write entire ref name (default: only up to 1st space)

Colorspace

  --snpphred <int>   Phred penalty for SNP when decoding colorspace (def: 30)
     or
  --snpfrac <dec>    approx. fraction of SNP bases (e.g. 0.001); sets --snpphred
  --col-cseq         print aligned colorspace seqs as colors, not decoded bases
  --col-cqual        print original colorspace quals, not decoded quals
  --col-keepends     keep nucleotides at extreme ends of decoded alignment

SAM

  -S/--sam           write hits in SAM format
  --mapq <int>       default mapping quality (MAPQ) to print for SAM alignments
  --sam-nohead       supppress header lines (starting with @) for SAM output
  --sam-nosq         supppress @SQ header lines for SAM output
  --sam-RG <text>    add <text> (usually "lab=value") to @RG line of SAM header

Performance

  -o/--offrate <int> override offrate of index; must be >= index's offrate
  -p/--threads <int> number of alignment threads to launch (default: 1)
  --mm               use memory-mapped I/O for index; many 'bowtie's can share
  --shmem            use shared mem for index; many 'bowtie's can share

Other

  --seed <int>       seed for random number generator
  --verbose          verbose output (for debugging)
  --version          print version information and quit
  -h/--help          print this usage message
1)
Note: Make sure the line:
export MODULEPATH=/opt/gridware/bioinformatics/modules:$MODULEPATH
exists in your ~/.profile file
/app/dokuwiki/data/attic/howto/bioinformatics/bowtie.1385119065.txt.gz · Last modified: 2021/12/09 16:42 (external edit)