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author | crs4 |
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date | Tue, 18 Mar 2014 07:49:22 -0400 |
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# -*- coding: utf-8 -*- #!/usr/bin/env python ## yufei.luo@gustave.roussy 22/07/2013 ## Copyright © 2014 CRS4 Srl. http://www.crs4.it/ ## Modified by: ## Nicola Soranzo <nicola.soranzo@crs4.it> """ 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_mem.py [options] See below for options """ import optparse, os, shutil, subprocess, sys, tempfile 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_mem.py: Map (long length) reads against a reference genome with BWA-MEM." parser = optparse.OptionParser(description=descr) parser.add_option( '-t', '--threads', default=1, help='The number of threads to use [1]' ) parser.add_option( '--ref', help='The reference genome to use or index' ) parser.add_option( '-f', '--fastq', help='The (forward) fastq file to use for the mapping' ) parser.add_option( '-F', '--rfastq', help='The reverse fastq file to use for mapping if paired-end data' ) parser.add_option( '-u', '--output', help='The file to save the output (SAM format)' ) parser.add_option( '-g', '--genAlignType', help='The type of pairing (single or paired)' ) parser.add_option( '--params', help='Parameter setting to use (pre_set or full)' ) parser.add_option( '-s', '--fileSource', help='Whether to use a previously indexed reference sequence or one form history (indexed or history)' ) parser.add_option( '-D', '--dbkey', help='Dbkey for reference genome' ) parser.add_option( '-k', '--minEditDistSeed', default=19, type=int, help='Minimum edit distance to the seed [19]' ) parser.add_option( '-w', '--bandWidth', default=100, type=int, help='Band width for banded alignment [100]' ) parser.add_option( '-d', '--offDiagonal', default=100, type=int, help='off-diagonal X-dropoff [100]' ) parser.add_option( '-r', '--internalSeeds', default=1.5, type=float, help='look for internal seeds inside a seed longer than {-k} * FLOAT [1.5]' ) parser.add_option( '-c', '--seedsOccurrence', default=10000, type=int, help='skip seeds with more than INT occurrences [10000]' ) parser.add_option( '-S', '--mateRescue', default=False, help='skip mate rescue' ) parser.add_option( '-P', '--skipPairing', default=False, help='skpe pairing, mate rescue performed unless -S also in use' ) parser.add_option( '-A', '--seqMatch', default=1, type=int, help='score of a sequence match' ) parser.add_option( '-B', '--mismatch', default=4, type=int, help='penalty for a mismatch' ) parser.add_option( '-O', '--gapOpen', default=6, type=int, help='gap open penalty' ) parser.add_option( '-E', '--gapExtension', default=None, help='gap extension penalty; a gap of size k cost {-O} + {-E}*k [1]' ) parser.add_option( '-L', '--clipping', default=5, type=int, help='penalty for clipping [5]' ) parser.add_option( '-U', '--unpairedReadpair', default=17, type=int, help='penalty for an unpaired read pair [17]' ) parser.add_option( '-p', '--interPairEnd', default=False, help='first query file consists of interleaved paired-end sequences' ) parser.add_option( '--rgid', help='Read group identifier' ) parser.add_option( '--rgsm', help='Sample' ) parser.add_option( '--rgpl', choices=[ 'CAPILLARY', 'LS454', 'ILLUMINA', 'SOLID', 'HELICOS', 'IONTORRENT', 'PACBIO' ], help='Platform/technology used to produce the reads' ) parser.add_option( '--rglb', help='Library name' ) parser.add_option( '--rgpu', help='Platform unit (e.g. flowcell-barcode.lane for Illumina or slide for SOLiD)' ) parser.add_option( '--rgcn', help='Sequencing center that produced the read' ) parser.add_option( '--rgds', help='Description' ) parser.add_option( '--rgdt', help='Date that run was produced (ISO8601 format date or date/time, like YYYY-MM-DD)' ) parser.add_option( '--rgfo', help='Flow order' ) parser.add_option( '--rgks', help='The array of nucleotide bases that correspond to the key sequence of each read' ) parser.add_option( '--rgpg', help='Programs used for processing the read group' ) parser.add_option( '--rgpi', help='Predicted median insert size' ) parser.add_option( '-T', '--minScore', default=30, type=int, help='minimum score to output [30]' ) parser.add_option( '-M', '--mark', default=False, help='mark shorter split hits as secondary (for Picard/GATK compatibility)' ) parser.add_option( '-H', '--suppressHeader', dest='suppressHeader', help='Suppress header' ) (options, args) = parser.parse_args() if len(args) > 0: parser.error('Wrong number of arguments') # 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' ) fastq = options.fastq if options.rfastq: rfastq = options.rfastq # make temp directory for placement of indices tmp_index_dir = tempfile.mkdtemp() tmp_dir = tempfile.mkdtemp() # index if necessary if options.fileSource == 'history': ref_file = tempfile.NamedTemporaryFile( dir=tmp_index_dir ) ref_file_name = ref_file.name ref_file.close() os.symlink( options.ref, ref_file_name ) # determine which indexing algorithm to use, based on size try: size = os.stat( options.ref ).st_size if size <= 2**30: indexingAlg = 'is' else: indexingAlg = 'bwtsw' except: indexingAlg = 'is' 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 = options.ref # if options.illumina13qual: # illumina_quals = "-I" # else: # illumina_quals = "" # set up aligning and generate aligning command args start_cmds = '-t %s ' % options.threads if options.params == 'pre_set': # aligning_cmds = '-t %s %s' % ( options.threads, illumina_quals ) #start_cmds = '-t %s ' % options.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 ' % (options.minEditDistSeed, options.bandWidth, options.offDiagonal, options.internalSeeds, options.seedsOccurrence, options.seqMatch, options.mismatch, options.gapOpen, options.clipping, options.unpairedReadpair, options.minScore) if options.mateRescue: end_cmds += '-S ' if options.skipPairing: end_cmds += '-P ' else: if options.skipPairing: print "Option Error and will not be considered, you should also choose 'skip mate rescue -S' option! " if options.gapExtension != None: end_cmds += '-E %s ' % options.gapExtension if options.rgid: if not options.rglb or not options.rgpl or not options.rgsm or not options.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' % ( options.rgid, options.rglb, options.rgpl, options.rgsm ) readGroup = '@RG\tID:%s\tLB:%s\tPL:%s\tSM:%s' % ( options.rgid, options.rglb, options.rgpl, options.rgsm ) if options.rgpu: readGroup += '\tPU:%s' % options.rgpu if options.rgcn: readGroup += '\tCN:%s' % options.rgcn if options.rgds: readGroup += '\tDS:%s' % options.rgds if options.rgdt: readGroup += '\tDT:%s' % options.rgdt if options.rgfo: readGroup += '\tFO:%s' % options.rgfo if options.rgks: readGroup += '\tKS:%s' % options.rgks if options.rgpg: readGroup += '\tPG:%s' % options.rgpg if options.rgpi: readGroup += '\tPI:%s' % options.rgpi end_cmds += ' -R "%s" ' % readGroup if options.interPairEnd: end_cmds += '-p %s ' % options.interPairEnd if options.mark: end_cmds += '-M ' if options.genAlignType == 'paired': cmd = 'bwa mem %s %s %s %s %s > %s' % ( start_cmds, ref_file_name, fastq, rfastq, end_cmds, options.output ) else: cmd = 'bwa mem %s %s %s %s > %s' % ( start_cmds, ref_file_name, fastq, end_cmds, options.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 ) # remove header if necessary if options.suppressHeader == 'true': tmp_out = tempfile.NamedTemporaryFile( dir=tmp_dir) tmp_out_name = tmp_out.name tmp_out.close() try: shutil.move( options.output, tmp_out_name ) except Exception, e: raise Exception, 'Error moving output file before removing headers. ' + str( e ) fout = file( options.output, 'w' ) for line in file( tmp_out.name, 'r' ): if not ( line.startswith( '@HD' ) or line.startswith( '@SQ' ) or line.startswith( '@RG' ) or line.startswith( '@PG' ) or line.startswith( '@CO' ) ): fout.write( line ) fout.close() # check that there are results in the output file if os.path.getsize( options.output ) > 0: sys.stdout.write( 'BWA run on %s-end data' % options.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__()