Mercurial > repos > yufei-luo > bwa_0_7_5
view bwa_0_7_5/bwa_0_7_5.py @ 4:22449c3a0b7f draft default tip
debug when use 'history' input fasta as genome reference
author | yufei-luo |
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date | Tue, 10 Dec 2013 08:48:23 -0500 |
parents | 8409cff2d740 |
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#!/usr/bin/env python ## yufei.luo@gustave.roussy 22/07/2013 """ Runs BWA on single-end or paired-end data. Produces a SAM file containing the mappings. Works with BWA version 0.7.5. NOTICE: In this wrapper, we only use 'mem' for mapping step. usage: bwa_0_7_5.py [args] See below for args """ import optparse, os, shutil, subprocess, sys, tempfile import argparse def stop_err( msg ): sys.stderr.write( '%s\n' % msg ) sys.exit() def check_is_double_encoded( fastq ): # check that first read is bases, not one base followed by numbers bases = [ 'A', 'C', 'G', 'T', 'a', 'c', 'g', 't', 'N' ] nums = [ '0', '1', '2', '3' ] for line in file( fastq, 'rb'): if not line.strip() or line.startswith( '@' ): continue if len( [ b for b in line.strip() if b in nums ] ) > 0: return False elif line.strip()[0] in bases and len( [ b for b in line.strip() if b in bases ] ) == len( line.strip() ): return True else: raise Exception, 'First line in first read does not appear to be a valid FASTQ read in either base-space or color-space' raise Exception, 'There is no non-comment and non-blank line in your FASTQ file' def __main__(): descr = "bwa_0_7_5.py: version 1.0. Map the reads(long length) against the genome reference with BWA MEM. \n" descr += "Usage: BWA mem -t thread -R groupInfo refSequence read.R1.fastq (read.R2.fastq) > out.sam" parser = argparse.ArgumentParser(description=descr) parser.add_argument( '-t', '--threads', default=1, help='The number of threads to use [1]' ) parser.add_argument( '--color-space', default=False, help='If the input files are SOLiD format' ) parser.add_argument( '--ref', help='The reference genome to use or index' ) parser.add_argument( '-f', '--fastq', help='The (forward) fastq file to use for the mapping' ) parser.add_argument( '-F', '--rfastq', help='The reverse fastq file to use for mapping if paired-end data' ) parser.add_argument( '-u', '--output', help='The file to save the output (SAM format)' ) parser.add_argument( '-g', '--genAlignType', help='The type of pairing (single or paired)' ) parser.add_argument( '--params', help='Parameter setting to use (pre_set or full)' ) parser.add_argument( '-s', '--fileSource', help='Whether to use a previously indexed reference sequence or one form history (indexed or history)' ) parser.add_argument( '-D', '--dbkey', help='Dbkey for reference genome' ) parser.add_argument( '-k', '--minEditDistSeed', default=19, type=int, help='Minimum edit distance to the seed [19]' ) parser.add_argument( '-w', '--bandWidth', default=100, type=int, help='Band width for banded alignment [100]' ) parser.add_argument( '-d', '--offDiagonal', default=100, type=int, help='off-diagonal X-dropoff [100]' ) parser.add_argument( '-r', '--internalSeeds', default=1.5, type=float, help='look for internal seeds inside a seed longer than {-k} * FLOAT [1.5]' ) parser.add_argument( '-c', '--seedsOccurrence', default=10000, type=int, help='skip seeds with more than INT occurrences [10000]' ) parser.add_argument( '-S', '--mateRescue', default=False, help='skip mate rescue' ) parser.add_argument( '-P', '--skipPairing', default=False, help='skpe pairing, mate rescue performed unless -S also in use' ) parser.add_argument( '-A', '--seqMatch', default=1, type=int, help='score of a sequence match' ) parser.add_argument( '-B', '--mismatch', default=4,type=int, help='penalty for a mismatch' ) parser.add_argument( '-O', '--gapOpen', default=6, type=int, help='gap open penalty' ) parser.add_argument( '-E', '--gapExtension', default=None, help='gap extension penalty; a gap of size k cost {-O} + {-E}*k [1]' ) parser.add_argument( '-L', '--clipping', default=5, type=int, help='penalty for clipping [5]' ) parser.add_argument( '-U', '--unpairedReadpair', default=17, type=int, help='penalty for an unpaired read pair [17]' ) parser.add_argument( '-p', '--interPairEnd', default=False, help='first query file consists of interleaved paired-end sequences' ) parser.add_argument( '--rgid', help='Read group identifier' ) parser.add_argument( '--rgsm', help='Sample' ) parser.add_argument( '--rgpl', choices=[ 'CAPILLARY', 'LS454', 'ILLUMINA', 'SOLID', 'HELICOS', 'IONTORRENT', 'PACBIO' ], help='Platform/technology used to produce the reads' ) parser.add_argument( '--rglb', help='Library name' ) parser.add_argument( '--rgpu', help='Platform unit (e.g. flowcell-barcode.lane for Illumina or slide for SOLiD)' ) parser.add_argument( '--rgcn', help='Sequencing center that produced the read' ) parser.add_argument( '--rgds', help='Description' ) parser.add_argument( '--rgdt', help='Date that run was produced (ISO8601 format date or date/time, like YYYY-MM-DD)' ) parser.add_argument( '--rgfo', help='Flow order' ) parser.add_argument( '--rgks', help='The array of nucleotide bases that correspond to the key sequence of each read' ) parser.add_argument( '--rgpg', help='Programs used for processing the read group' ) parser.add_argument( '--rgpi', help='Predicted median insert size' ) parser.add_argument( '-T', '--minScore', default=30, type=int, help='minimum score to output [30]' ) parser.add_argument( '-M', '--mark', default=False, help='mark shorter split hits as secondary (for Picard/GATK compatibility)' ) args = parser.parse_args() # output version # of tool try: tmp = tempfile.NamedTemporaryFile().name tmp_stdout = open( tmp, 'wb' ) proc = subprocess.Popen( args='bwa 2>&1', shell=True, stdout=tmp_stdout ) tmp_stdout.close() returncode = proc.wait() stdout = None for line in open( tmp_stdout.name, 'rb' ): if line.lower().find( 'version' ) >= 0: stdout = line.strip() break if stdout: sys.stdout.write( 'BWA %s\n' % stdout ) else: raise Exception except: sys.stdout.write( 'Could not determine BWA version\n' ) # check for color space fastq that's not double-encoded and exit if appropriate # if args.color_space: # if not check_is_double_encoded( args.fastq ): # stop_err( 'Your file must be double-encoded (it must be converted from "numbers" to "bases"). See the help section for details' ) # if args.genAlignType == 'paired': # if not check_is_double_encoded( args.rfastq ): # stop_err( 'Your reverse reads file must also be double-encoded (it must be converted from "numbers" to "bases"). See the help section for details' ) fastq = args.fastq if args.rfastq: rfastq = args.rfastq # set color space variable # if args.color_space: # color_space = '-c' # else: # color_space = '' # make temp directory for placement of indices tmp_index_dir = tempfile.mkdtemp() tmp_dir = tempfile.mkdtemp() # index if necessary # if args.fileSource == 'history' and not args.do_not_build_index: if args.fileSource == 'history' : ref_file = tempfile.NamedTemporaryFile( dir=tmp_index_dir ) ref_file_name = ref_file.name ref_file.close() os.symlink( args.ref, ref_file_name ) # determine which indexing algorithm to use, based on size try: size = os.stat( args.ref ).st_size if size <= 2**30: indexingAlg = 'is' else: indexingAlg = 'bwtsw' except: indexingAlg = 'is' #indexing_cmds = '%s -a %s' % ( color_space, indexingAlg ) indexing_cmds = '-a %s' % indexingAlg cmd1 = 'bwa index %s %s' % ( indexing_cmds, ref_file_name ) try: tmp = tempfile.NamedTemporaryFile( dir=tmp_index_dir ).name tmp_stderr = open( tmp, 'wb' ) proc = subprocess.Popen( args=cmd1, shell=True, cwd=tmp_index_dir, stderr=tmp_stderr.fileno() ) returncode = proc.wait() tmp_stderr.close() # get stderr, allowing for case where it's very large tmp_stderr = open( tmp, 'rb' ) stderr = '' buffsize = 1048576 try: while True: stderr += tmp_stderr.read( buffsize ) if not stderr or len( stderr ) % buffsize != 0: break except OverflowError: pass tmp_stderr.close() if returncode != 0: raise Exception, stderr except Exception, e: # clean up temp dirs if os.path.exists( tmp_index_dir ): shutil.rmtree( tmp_index_dir ) if os.path.exists( tmp_dir ): shutil.rmtree( tmp_dir ) stop_err( 'Error indexing reference sequence. ' + str( e ) ) else: ref_file_name = args.ref # if args.illumina13qual: # illumina_quals = "-I" # else: # illumina_quals = "" # set up aligning and generate aligning command args start_cmds = '-t %s ' % args.threads if args.params == 'pre_set': # aligning_cmds = '-t %s %s %s' % ( args.threads, color_space, illumina_quals ) #start_cmds = '-t %s ' % args.threads end_cmds = ' ' print start_cmds, end_cmds else: end_cmds = '-k %s -w %s -d %s -r %s -c %s -A %s -B %s -O %s -L %s -U %s -T %s ' % (args.minEditDistSeed, args.bandWidth, args.offDiagonal, args.internalSeeds, args.seedsOccurrence, args.seqMatch, args.mismatch, args.gapOpen, args.clipping, args.unpairedReadpair, args.minScore) if args.mateRescue: end_cmds += '-S ' if args.skipPairing: end_cmds += '-P ' else: if args.skipPairing: print "Option Error and will not be considered, you should also choose 'skip mate rescue -S' option! " if args.gapExtension != None: end_cmds += '-E %s ' % args.gapExtension if args.rgid: if not args.rglb or not args.rgpl or not args.rgsm or not args.rglb: stop_err( 'If you want to specify read groups, you must include the ID, LB, PL, and SM tags.' ) # readGroup = '@RG\tID:%s\tLB:%s\tPL:%s\tSM:%s' % ( args.rgid, args.rglb, args.rgpl, args.rgsm ) readGroup = '@RG\tID:%s\tLB:%s\tPL:%s\tSM:%s' % ( args.rgid, args.rglb, args.rgpl, args.rgsm ) if args.rgpu: readGroup += '\tPU:%s' % args.rgpu if args.rgcn: readGroup += '\tCN:%s' % args.rgcn if args.rgds: readGroup += '\tDS:%s' % args.rgds if args.rgdt: readGroup += '\tDT:%s' % args.rgdt if args.rgfo: readGroup += '\tFO:%s' % args.rgfo if args.rgks: readGroup += '\tKS:%s' % args.rgks if args.rgpg: readGroup += '\tPG:%s' % args.rgpg if args.rgpi: readGroup += '\tPI:%s' % args.rgpi end_cmds += ' -R "%s" ' % readGroup if args.interPairEnd: end_cmds += '-p %s ' % args.interPairEnd if args.mark: end_cmds += '-M ' if args.genAlignType == 'paired': cmd = 'bwa mem %s %s %s %s %s > %s' % ( start_cmds, ref_file_name, fastq, rfastq, end_cmds, args.output ) else: cmd = 'bwa mem %s %s %s %s > %s' % ( start_cmds, ref_file_name, fastq, end_cmds, args.output ) # perform alignments buffsize = 1048576 try: # need to nest try-except in try-finally to handle 2.4 try: try: tmp = tempfile.NamedTemporaryFile( dir=tmp_dir ).name tmp_stderr = open( tmp, 'wb' ) print "The cmd is %s" % cmd proc = subprocess.Popen( args=cmd, shell=True, cwd=tmp_dir, stderr=tmp_stderr.fileno() ) returncode = proc.wait() tmp_stderr.close() # get stderr, allowing for case where it's very large tmp_stderr = open( tmp, 'rb' ) stderr = '' try: while True: stderr += tmp_stderr.read( buffsize ) if not stderr or len( stderr ) % buffsize != 0: break except OverflowError: pass tmp_stderr.close() if returncode != 0: raise Exception, stderr except Exception, e: raise Exception, 'Error generating alignments. ' + str( e ) # check that there are results in the output file if os.path.getsize( args.output ) > 0: sys.stdout.write( 'BWA run on %s-end data' % args.genAlignType ) else: raise Exception, 'The output file is empty. You may simply have no matches, or there may be an error with your input file or settings.' except Exception, e: stop_err( 'The alignment failed.\n' + str( e ) ) finally: # clean up temp dir if os.path.exists( tmp_index_dir ): shutil.rmtree( tmp_index_dir ) if os.path.exists( tmp_dir ): shutil.rmtree( tmp_dir ) if __name__=="__main__": __main__()