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|
#!/bin/bash
usage(){
echo "
BBMap
Written by Brian Bushnell, from Dec. 2010 - present
Last modified September 15, 2022
Description: Fast and accurate splice-aware read aligner.
Please read bbmap/docs/guides/BBMapGuide.txt for more information.
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.
If indexing and mapping are done in two steps, K should
be specified each time.
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.
tipsearch=100 Look this far for read-end deletions with anchors
shorter than K, using brute force.
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.
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.
deterministic=f Run in deterministic mode. In this case it is good
to set averagepairdist. BBMap is deterministic
without this flag if using single-ended reads,
or run singlethreaded.
bandwidthratio=0 (bwr) If above zero, restrict alignment band to this
fraction of read length. Faster but less accurate.
bandwidth=0 (bw) Set the bandwidth directly.
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.
ignorefrequentkmers=t (ifk) Discard low-information kmers that occur often.
excludefraction=0.03 (ef) Fraction of kmers to ignore. For example, 0.03
will ignore the most common 3% of kmers.
greedy=t Use a greedy algorithm to discard the least-useful
kmers on a per-read basis.
kfilter=0 If positive, potential mapping sites must have at
least this many consecutive exact matches.
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) Do not map 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 '.').
renamebyinsert=f Renames reads based on their mapped insert size.
Bloom-Filtering Parameters (bloomfilter.sh is the standalone version).
bloom=f Use a Bloom filter to ignore reads not sharing kmers
with the reference. This uses more memory, but speeds
mapping when most reads don't match the reference.
bloomhashes=2 Number of hash functions.
bloomminhits=3 Number of consecutive hits to be considered matched.
bloomk=31 Bloom filter kmer length.
bloomserial=t Use the serialized Bloom filter for greater loading
speed, if available. If not, generate and write one.
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.
nfilter=-1 Ban alignments with more than this many ns. This
includes nocall, noref, and off scaffold ends.
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=
If you set a readgroup flag to the word 'filename',
e.g. rgid=filename, the input file name will be used.
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.
gcpairs=t Use average GC of paired reads.
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.
covk=0 If positive, calculate kmer coverage statistics.
Java Parameters:
-Xmx This will set Java's memory usage,
overriding autodetection.
-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.
-eoom This flag will cause the process to exit if an
out-of-memory exception occurs. Requires Java 8u92+.
-da Disable assertions.
Please contact Brian Bushnell at bbushnell@lbl.gov if you encounter
any problems, or post at: http://seqanswers.com/forums/showthread.php?t=41057
"
}
#This block allows symlinked shellscripts to correctly set classpath.
pushd . > /dev/null
DIR="${BASH_SOURCE[0]}"
while [ -h "$DIR" ]; do
cd "$(dirname "$DIR")"
DIR="$(readlink "$(basename "$DIR")")"
done
cd "$(dirname "$DIR")"
DIR="$(pwd)/"
popd > /dev/null
#DIR="$( cd "$( dirname "${BASH_SOURCE[0]}" )" && pwd )/"
CP="$DIR""current/"
JNI="-Djava.library.path=""$DIR""jni/"
JNI=""
z="-Xmx1g"
z2="-Xms1g"
set=0
if [ -z "$1" ] || [[ $1 == -h ]] || [[ $1 == --help ]]; then
usage
exit
fi
calcXmx () {
source "$DIR""/calcmem.sh"
setEnvironment
parseXmx "$@"
if [[ $set == 1 ]]; then
return
fi
freeRam 3200m 84
z="-Xmx${RAM}m"
z2="-Xms${RAM}m"
}
calcXmx "$@"
bbmap() {
local CMD="java $EA $EOOM $z $z2 $JNI -cp $CP align2.BBMap build=1 overwrite=true fastareadlen=500 $@"
echo $CMD >&2
eval $CMD
}
bbmap "$@"
|
