bowtie2 - Online in the Cloud

PROGRAM:

NAME

bowtie2 - wrapper for bowtie2-align-*

DESCRIPTION

Bowtie 2 version 2.2.6 by Ben Langmead ([email protected], www.cs.jhu.edu/~langmea)

USAGE

bowtie2 [options]* -x <bt2-idx> {-1 <m1> -2 <m2> | -U <r>} [-S <sam>]

<bt2-idx>
Index filename prefix (minus trailing .X.bt2). NOTE: Bowtie 1 and Bowtie 2 indexes
are not compatible.

<m1> Files with #1 mates, paired with files in <m2>. Could be gzip'ed (extension: .gz)
or bzip2'ed (extension: .bz2).

<m2> Files with #2 mates, paired with files in <m1>. Could be gzip'ed (extension: .gz)
or bzip2'ed (extension: .bz2).

<r> Files with unpaired reads. Could be gzip'ed (extension: .gz) or bzip2'ed
(extension: .bz2).

<sam> File for SAM output (default: stdout)

<m1>, <m2>, <r> can be comma-separated lists (no whitespace) and can be specified
many times. E.g. '-U file1.fq,file2.fq -U file3.fq'.

OPTIONS (defaults in parentheses)

Input:
-q query input files are FASTQ .fq/.fastq (default)

--qseq query input files are in Illumina's qseq format

-f query input files are (multi-)FASTA .fa/.mfa

-r query input files are raw one-sequence-per-line

-c <m1>, <m2>, <r> are sequences themselves, not files

-s/--skip <int>
skip the first <int> reads/pairs in the input (none)

-u/--upto <int>
stop after first <int> reads/pairs (no limit)

-5/--trim5 <int>
trim <int> bases from 5'/left end of reads (0)

-3/--trim3 <int>
trim <int> bases from 3'/right end of reads (0)

--phred33
qualities are Phred+33 (default)

--phred64
qualities are Phred+64

--int-quals
qualities encoded as space-delimited integers

Presets:
Same as:

For --end-to-end:

--very-fast -D 5 -R 1 -N 0 -L 22 -i S,0,2.50

--fast -D 10 -R 2 -N 0 -L 22 -i S,0,2.50

--sensitive -D 15 -R 2 -N 0 -L 22 -i S,1,1.15 (default)

--very-sensitive -D 20 -R 3 -N 0 -L 20 -i S,1,0.50

For --local:

--very-fast-local -D 5 -R 1 -N 0 -L 25 -i S,1,2.00

--fast-local -D 10 -R 2 -N 0 -L 22 -i S,1,1.75

--sensitive-local -D 15 -R 2 -N 0 -L 20 -i S,1,0.75 (default)

--very-sensitive-local -D 20 -R 3 -N 0 -L 20 -i S,1,0.50

Alignment:
-N <int>
max # mismatches in seed alignment; can be 0 or 1 (0)

-L <int>
length of seed substrings; must be >3, <32 (22)

-i <func>
interval between seed substrings w/r/t read len (S,1,1.15)

--n-ceil <func>
func for max # non-A/C/G/Ts permitted in aln (L,0,0.15)

--dpad <int>
include <int> extra ref chars on sides of DP table (15)

--gbar <int>
disallow gaps within <int> nucs of read extremes (4)

--ignore-quals
treat all quality values as 30 on Phred scale (off)

--nofw do not align forward (original) version of read (off)

--norc do not align reverse-complement version of read (off)

--no-1mm-upfront
do not allow 1 mismatch alignments before attempting to scan for the optimal seeded
alignments

--end-to-end
entire read must align; no clipping (on)

OR

--local
local alignment; ends might be soft clipped (off)

Scoring:
--ma <int>
match bonus (0 for --end-to-end, 2 for --local)

--mp <int>
max penalty for mismatch; lower qual = lower penalty (6)

--np <int>
penalty for non-A/C/G/Ts in read/ref (1)

--rdg <int>,<int>
read gap open, extend penalties (5,3)

--rfg <int>,<int>
reference gap open, extend penalties (5,3)

--score-min <func> min acceptable alignment score w/r/t read length
(G,20,8 for local, L,-0.6,-0.6 for end-to-end)

Reporting:
(default)
look for multiple alignments, report best, with MAPQ

OR

-k <int>
report up to <int> alns per read; MAPQ not meaningful

OR

-a/--all
report all alignments; very slow, MAPQ not meaningful

Effort:
-D <int>
give up extending after <int> failed extends in a row (15)

-R <int>
for reads w/ repetitive seeds, try <int> sets of seeds (2)

Paired-end:

-I/--minins <int>
minimum fragment length (0)

-X/--maxins <int>
maximum fragment length (500)

--fr/--rf/--ff -1, -2 mates align fw/rev, rev/fw, fw/fw (--fr)

--no-mixed
suppress unpaired alignments for paired reads

--no-discordant
suppress discordant alignments for paired reads

--no-dovetail
not concordant when mates extend past each other

--no-contain
not concordant when one mate alignment contains other

--no-overlap
not concordant when mates overlap at all

Output:
-t/--time
print wall-clock time taken by search phases

--un <path>
write unpaired reads that didn't align to <path>

--al <path>
write unpaired reads that aligned at least once to <path>

--un-conc <path>
write pairs that didn't align concordantly to <path>

--al-conc <path>
write pairs that aligned concordantly at least once to <path>

(Note: for --un, --al, --un-conc, or --al-conc, add '-gz' to the option name, e.g.
--un-gz <path>, to gzip compress output, or add '-bz2' to bzip2 compress output.)
--quiet print nothing to stderr except serious errors
--met-file <path> send metrics to file at <path> (off)
--met-stderr send metrics to stderr (off)
--met <int> report internal counters & metrics every <int> secs (1)
--no-unal supppress SAM records for unaligned reads
--no-head supppress header lines, i.e. lines starting with @
--no-sq supppress @SQ header lines
--rg-id <text> set read group id, reflected in @RG line and RG:Z: opt field
--rg <text> add <text> ("lab:value") to @RG line of SAM header.

Note: @RG line only printed when --rg-id is set.

--omit-sec-seq
put '*' in SEQ and QUAL fields for secondary alignments.

Performance:
-p/--threads <int> number of alignment threads to launch (1)

--reorder
force SAM output order to match order of input reads

--mm use memory-mapped I/O for index; many 'bowtie's can share

Other:
--qc-filter
filter out reads that are bad according to QSEQ filter

--seed <int>
seed for random number generator (0)

--non-deterministic seed rand. gen. arbitrarily instead of using read attributes

--version
print version information and quit

-h/--help
print this usage message

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