bowtie2-align1
Ultrafast and memory-efficient tool for aligning sequencing reads to long reference sequences
Overview
Bowtie 2 version 2.0.0-beta6 by Ben Langmead ([email protected]) Usage:
-
bowtie2-align [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>.
- <m2>
-
Files with #2 mates, paired with files in <m1>.
- <r>
-
Files with unpaired reads.
- <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
-Ufile3.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,1,1.25 (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,1,1.25)
-
--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)
-
--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:
-
-M <int> -
look for up to <int>+1 alns; report best, with MAPQ (5 for
--end-to-end, 2 for--local)
-
OR
-
-k <int> -
report up to <int> alns per read; MAPQ not meaningful (off)
-
OR
-a/--all-
report all alignments; very slow (off)
-
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
-
--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-idis set.
-
Performance:
-o/--offrate <int> override offrate of index; must be >= index's offrate
-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)
-
--version -
print version information and quit
-h/--help-
print this usage message
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