BBMap

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Category

Bioinformatics

Program On

zcluster

Version

35.66

Author / Distributor

BBMap

Description

"Short read aligner for DNA and RNA-seq data and others..." More details at BBMap

Running Program

Also refer to Running Jobs on zcluster

Latest version /usr/local/bbmap/latest points to /usr/local/bbmap/35.66

Example of a shell script sub.sh to run on at the batch queue:

#!/bin/bash
cd working_directory
time  /usr/local/bbmap/latest/bbmap.sh [options] 

Example of job submission to the queue:

qsub -q queuename sub.sh

Documentation

 
/usr/local/bbmap/latest/bbmap.sh

BBMap v35.x
Written by Brian Bushnell, from Dec. 2010 - present
Last modified October 23, 2015

Description:  Fast and accurate splice-aware read aligner.

To index:     bbmap.sh ref=<reference fasta>
To map:       bbmap.sh in=<reads> out=<output sam>
To map without writing an index:
    bbmap.sh ref=<reference fasta> in=<reads> out=<output sam> nodisk

in=stdin will accept reads from standard in, and out=stdout will write to 
standard out, but file extensions are still needed to specify the format of the 
input and output files e.g. in=stdin.fa.gz will read gzipped fasta from 
standard in; out=stdout.sam.gz will write gzipped sam.

Indexing Parameters (required when building the index):

nodisk=f                Set to true to build index in memory and write nothing 
                        to disk except output.
ref=<file>              Specify the reference sequence.  Only do this ONCE, 
                        when building the index (unless using 'nodisk').
build=1                 If multiple references are indexed in the same directory,
                        each needs a unique numeric ID (unless using 'nodisk').
k=13                    Kmer length, range 8-15.  Longer is faster but uses 
                        more memory.  Shorter is more sensitive.
path=<.>                Specify the location to write the index, if you don't 
                        want it in the current working directory.
usemodulo=f             Throw away ~80% of kmers based on remainder modulo a 
                        number (reduces RAM by 50% and sensitivity slightly).
                        Should be enabled both when building the index AND 
                        when mapping.
rebuild=f               Force a rebuild of the index (ref= should be set).

Input Parameters:

build=1                 Designate index to use.  Corresponds to the number 
                        specified when building the index.
in=<file>               Primary reads input; required parameter.
in2=<file>              For paired reads in two files.
interleaved=auto        True forces paired/interleaved input; false forces 
                        single-ended mapping. If not specified, interleaved 
                        status will be autodetected from read names.
fastareadlen=500        Break up FASTA reads longer than this.  Max is 500 for
                        BBMap and 6000 for BBMapPacBio.  Only works for FASTA
                        input (use 'maxlen' for FASTQ input).  The default for
                        bbmap.sh is 500, and for mapPacBio.sh is 6000.
unpigz=f                Spawn a pigz (parallel gzip) process for faster 
                        decompression than using Java.  
                        Requires pigz to be installed.
touppercase=t           (tuc) Convert lowercase letters in reads to upper case 
                        (otherwise they will not match the reference).

Sampling Parameters:

reads=-1                Set to a positive number N to only process the first N
                        reads (or pairs), then quit.  -1 means use all reads.
samplerate=1            Set to a number from 0 to 1 to randomly select that
                        fraction of reads for mapping. 1 uses all reads.
skipreads=0             Set to a number N to skip the first N reads (or pairs), 
                        then map the rest.

Mapping Parameters:

fast=f                  This flag is a macro which sets other paramters to run 
                        faster, at reduced sensitivity.  Bad for RNA-seq.
slow=f                  This flag is a macro which sets other paramters to run 
                        slower, at greater sensitivity.  'vslow' is even slower.
maxindel=16000          Don't look for indels longer than this. Lower is faster.
                        Set to >=100k for RNAseq with long introns like mammals.
strictmaxindel=f        When enabled, do not allow indels longer than 'maxindel'.
                        By default these are not sought, but may be found anyway.
minid=0.76              Approximate minimum alignment identity to look for. 
                        Higher is faster and less sensitive.
minhits=1               Minimum number of seed hits required for candidate sites.
                        Higher is faster.
k=13                    Kmer length of index.  Lower is slower, more sensitive,
                        and uses slightly less RAM.  Range is 7 to 15.
local=f                 Set to true to use local, rather than global, alignments.
                        This will soft-clip ugly ends of poor alignments.
perfectmode=f           Allow only perfect mappings when set to true (very fast).
semiperfectmode=f       Allow only perfect and semiperfect (perfect except for 
                        N's in the reference) mappings.
threads=auto            (t) Set to number of threads desired.  By default, uses 
                        all cores available.
ambiguous=best          (ambig) Set behavior on ambiguously-mapped reads (with 
                        multiple top-scoring mapping locations).
                            best    (use the first best site)
                            toss    (consider unmapped)
                            random  (select one top-scoring site randomly)
                            all     (retain all top-scoring sites)
samestrandpairs=f       (ssp) Specify whether paired reads should map to the
                        same strand or opposite strands.
requirecorrectstrand=t  (rcs) Forbid pairing of reads without correct strand 
                        orientation.  Set to false for long-mate-pair libraries.
killbadpairs=f          (kbp) If a read pair is mapped with an inappropriate
                        insert size or orientation, the read with the lower  
                        mapping quality is marked unmapped.
pairedonly=f            (po) Treat unpaired reads as unmapped.  Thus they will 
                        be sent to 'outu' but not 'outm'.
rcomp=f                 Reverse complement both reads prior to mapping (for LMP
                        outward-facing libraries).
rcompmate=f             Reverse complement read2 prior to mapping.
pairlen=32000           Set max allowed distance between paired reads.  
                        (insert size)=(pairlen)+(read1 length)+(read2 length)
rescuedist=1200         Don't try to rescue paired reads if avg. insert size
                        greater than this.  Lower is faster.
rescuemismatches=32     Maximum mismatches allowed in a rescued read.  Lower
                        is faster.
averagepairdist=100     (apd) Initial average distance between paired reads.
                        Varies dynamically; does not need to be specified.
bandwidthratio=0        (bwr) If above zero, restrict alignment band to this 
                        fraction of read length.  Faster but less accurate.
usejni=f                (jni) Do alignments faster, in C code.  Requires 
                        compiling the C code; details are in /jni/README.txt.
maxsites2=800           Don't analyze (or print) more than this many alignments 
                        per read.
monitor=f               Kill this process if CPU usage drops to zero for
                        a long time.  monitor=600,0.01 would kill after 600
                        seconds under 1% usage.

Quality and Trimming Parameters:

qin=auto                Set to 33 or 64 to specify input quality value ASCII
                        offset. 33 is Sanger, 64 is old Solexa.
qout=auto               Set to 33 or 64 to specify output quality value ASCII 
                        offset (only if output format is fastq).
qtrim=f                 Quality-trim ends before mapping.  Options are: 
                        'f' (false), 'l' (left), 'r' (right), and 'lr' (both).
untrim=f                Undo trimming after mapping.  Untrimmed bases will be 
                        soft-clipped in cigar strings.
trimq=6                 Trim regions with average quality below this 
                        (phred algorithm).
mintrimlength=60        (mintl) Don't trim reads to be shorter than this.
fakefastaquality=-1     (ffq) Set to a positive number 1-50 to generate fake
                        quality strings for fasta input reads.
ignorebadquality=f      (ibq) Keep going, rather than crashing, if a read has 
                        out-of-range quality values.
usequality=t            Use quality scores when determining which read kmers
                        to use as seeds.
minaveragequality=0     (maq) Discard reads with average quality below this.
maqb=0                  If positive, calculate maq from this many initial bases.

Output Parameters:

out=<file>              Write all reads to this file.
outu=<file>             Write only unmapped reads to this file.  Does not 
                        include unmapped paired reads with a mapped mate.
outm=<file>             Write only mapped reads to this file.  Includes 
                        unmapped paired reads with a mapped mate.
mappedonly=f            If true, treats 'out' like 'outm'.
bamscript=<file>        (bs) Write a shell script to <file> that will turn 
                        the sam output into a sorted, indexed bam file.
ordered=f               Set to true to output reads in same order as input.  
                        Slower and uses more memory.
overwrite=f             (ow) Allow process to overwrite existing files.
secondary=f             Print secondary alignments.
sssr=0.95               (secondarysitescoreratio) Print only secondary alignments
                        with score of at least this fraction of primary.
ssao=f                  (secondarysiteasambiguousonly) Only print secondary 
                        alignments for ambiguously-mapped reads.
maxsites=5              Maximum number of total alignments to print per read.
                        Only relevant when secondary=t.
quickmatch=f            Generate cigar strings more quickly.
trimreaddescriptions=f  (trd) Truncate read and ref names at the first whitespace,
                        assuming that the remainder is a comment or description.
ziplevel=2              (zl) Compression level for zip or gzip output.
pigz=f                  Spawn a pigz (parallel gzip) process for faster 
                        compression than Java.  Requires pigz to be installed.
machineout=f            Set to true to output statistics in machine-friendly 
                        'key=value' format.
printunmappedcount=f    Print the total number of unmapped reads and bases.
                        If input is paired, the number will be of pairs
                        for which both reads are unmapped.
showprogress=0          If positive, print a '.' every X reads.
showprogress2=0         If positive, print the number of seconds since the
                        last progress update (instead of a '.').

Post-Filtering Parameters:

idfilter=0              Independant of minid; sets exact minimum identity 
                        allowed for alignments to be printed.  Range 0 to 1.
subfilter=-1            Ban alignments with more than this many substitutions.
insfilter=-1            Ban alignments with more than this many insertions.
delfilter=-1            Ban alignments with more than this many deletions.
indelfilter=-1          Ban alignments with more than this many indels.
editfilter=-1           Ban alignments with more than this many edits.
inslenfilter=-1         Ban alignments with an insertion longer than this.
dellenfilter=-1         Ban alignments with a deletion longer than this.

Sam flags and settings:

noheader=f              Disable generation of header lines.
sam=1.4                 Set to 1.4 to write Sam version 1.4 cigar strings, 
                        with = and X, or 1.3 to use M.
saa=t                   (secondaryalignmentasterisks) Use asterisks instead of
                        bases for sam secondary alignments.
cigar=t                 Set to 'f' to skip generation of cigar strings (faster).
keepnames=f             Keep original names of paired reads, rather than 
                        ensuring both reads have the same name.
intronlen=999999999     Set to a lower number like 10 to change 'D' to 'N' in 
                        cigar strings for deletions of at least that length.
rgid=                   Set readgroup ID.  All other readgroup fields 
                        can be set similarly, with the flag rgXX=
mdtag=f                 Write MD tags.
nhtag=f                 Write NH tags.
xmtag=f                 Write XM tags (may only work correctly with ambig=all).
amtag=f                 Write AM tags.
nmtag=f                 Write NM tags.
xstag=f                 Set to 'xs=fs', 'xs=ss', or 'xs=us' to write XS tags 
                        for RNAseq using firststrand, secondstrand, or 
                        unstranded libraries.  Needed by Cufflinks.  
                        JGI mainly uses 'firststrand'.
stoptag=f               Write a tag indicating read stop location, prefixed by YS:i:
lengthtag=f             Write a tag indicating (query,ref) alignment lengths, 
                        prefixed by YL:Z:
idtag=f                 Write a tag indicating percent identity, prefixed by YI:f:
inserttag=f             Write a tag indicating insert size, prefixed by X8:Z:
scoretag=f              Write a tag indicating BBMap's raw score, prefixed by YR:i:
timetag=f               Write a tag indicating this read's mapping time, prefixed by X0:i:
boundstag=f             Write a tag indicating whether either read in the pair
                        goes off the end of the reference, prefixed by XB:Z:
notags=f                Turn off all optional tags.

Histogram and statistics output parameters:

scafstats=<file>        Statistics on how many reads mapped to which scaffold.
refstats=<file>         Statistics on how many reads mapped to which reference
                        file; only for BBSplit.
sortscafs=t             Sort scaffolds or references by read count.
bhist=<file>            Base composition histogram by position.
qhist=<file>            Quality histogram by position.
aqhist=<file>           Histogram of average read quality.
bqhist=<file>           Quality histogram designed for box plots.
lhist=<file>            Read length histogram.
ihist=<file>            Write histogram of insert sizes (for paired reads).
ehist=<file>            Errors-per-read histogram.
qahist=<file>           Quality accuracy histogram of error rates versus 
                        quality score.
indelhist=<file>        Indel length histogram.
mhist=<file>            Histogram of match, sub, del, and ins rates by 
                        read location.
gchist=<file>           Read GC content histogram.
gcbins=100              Number gchist bins.  Set to 'auto' to use read length.
idhist=<file>           Histogram of read count versus percent identity.
idbins=100              Number idhist bins.  Set to 'auto' to use read length.
statsfile=stderr        Mapping statistics are printed here.

Coverage output parameters (these may reduce speed and use more RAM):

covstats=<file>         Per-scaffold coverage info.
rpkm=<file>             Per-scaffold RPKM/FPKM counts.
covhist=<file>          Histogram of # occurrences of each depth level.
basecov=<file>          Coverage per base location.
bincov=<file>           Print binned coverage per location (one line per X bases).
covbinsize=1000         Set the binsize for binned coverage output.
nzo=t                   Only print scaffolds with nonzero coverage.
twocolumn=f             Change to true to print only ID and Avg_fold instead of 
                        all 6 columns to the 'out=' file.
32bit=f                 Set to true if you need per-base coverage over 64k.
strandedcov=f           Track coverage for plus and minus strand independently.
startcov=f              Only track start positions of reads.
secondarycov=t          Include coverage of secondary alignments.
physcov=f               Calculate physical coverage for paired reads.
                        This includes the unsequenced bases.
delcoverage=t           (delcov) Count bases covered by deletions as covered.
                        True is faster than false.

Java Parameters:
-Xmx                    This will be passed to Java to set memory usage, 
                        overriding the program's automatic memory detection.
                        -Xmx20g will specify 20 gigs of RAM, and -Xmx800m 
                        will specify 800 megs.  The max is typically 85% of 
                        physical memory.  The human genome requires around 24g,
                        or 12g with the 'usemodulo' flag.  The index uses 
                        roughly 6 bytes per reference base.

This list is not complete.  For more information, please consult 
/usr/local/bbmap/latest/docs/readme.txt. 
Please contact Brian Bushnell at bbushnell@lbl.gov if you encounter 
any problems, or post at: http://seqanswers.com/forums/showthread.php?t=41057


Installation

Binary from BBMap

System

64-bit Linux