#!/usr/bin/env python """ Creates a pileup file from a bam file and a reference. usage: %prog [options] -p, --input1=p: bam file -o, --output1=o: Output pileup -R, --ref=R: Reference file type -n, --ownFile=n: User-supplied fasta reference file -d, --dbkey=d: dbkey of user-supplied file -x, --indexDir=x: Index directory -b, --bamIndex=b: BAM index file -s, --lastCol=s: Print the mapping quality as the last column -i, --indels=i: Only output lines containing indels -M, --mapCap=M: Cap mapping quality -c, --consensus=c: Call the consensus sequence using MAQ consensu model -T, --theta=T: Theta paramter (error dependency coefficient) -N, --hapNum=N: Number of haplotypes in sample -r, --fraction=r: Expected fraction of differences between a pair of haplotypes -I, --phredProb=I: Phred probability of an indel in sequencing/prep """ import os, shutil, subprocess, sys, tempfile from galaxy import eggs import pkg_resources; pkg_resources.require( "bx-python" ) from bx.cookbook import doc_optparse def stop_err( msg ): sys.stderr.write( '%s\n' % msg ) sys.exit() def check_seq_file( dbkey, GALAXY_DATA_INDEX_DIR ): seqFile = '%s/sam_fa_indices.loc' % GALAXY_DATA_INDEX_DIR seqPath = '' for line in open( seqFile ): line = line.rstrip( '\r\n' ) if line and not line.startswith( '#' ) and line.startswith( 'index' ): fields = line.split( '\t' ) if len( fields ) < 3: continue if fields[1] == dbkey: seqPath = fields[2].strip() break return seqPath def __main__(): #Parse Command Line options, args = doc_optparse.parse( __doc__ ) seqPath = check_seq_file( options.dbkey, options.indexDir ) #prepare file names tmpDir = tempfile.mkdtemp() tmpf0 = tempfile.NamedTemporaryFile( dir=tmpDir ) tmpf0_name = tmpf0.name tmpf0.close() tmpf0bam_name = '%s.bam' % tmpf0_name tmpf0bambai_name = '%s.bam.bai' % tmpf0_name tmpf1 = tempfile.NamedTemporaryFile( dir=tmpDir ) tmpf1_name = tmpf1.name tmpf1.close() tmpf1fai_name = '%s.fai' % tmpf1_name #link bam and bam index to working directory (can't move because need to leave original) os.symlink( options.input1, tmpf0bam_name ) os.symlink( options.bamIndex, tmpf0bambai_name ) #get parameters for pileup command if options.lastCol == 'yes': lastCol = '-s' else: lastCol = '' if options.indels == 'yes': indels = '-i' else: indels = '' opts = '%s %s -M %s' % ( lastCol, indels, options.mapCap ) if options.consensus == 'yes': opts += ' -c -T %s -N %s -r %s -I %s' % ( options.theta, options.hapNum, options.fraction, options.phredProb ) #prepare basic pileup command cmd = 'samtools pileup %s -f %s %s > %s' try: # have to nest try-except in try-finally to handle 2.4 try: #index reference if necessary and prepare pileup command if options.ref == 'indexed': if not os.path.exists( "%s.fai" % seqPath ): raise Exception, "No sequences are available for '%s', request them by reporting this error." % options.dbkey cmd = cmd % ( opts, seqPath, tmpf0bam_name, options.output1 ) elif options.ref == 'history': os.symlink( options.ownFile, tmpf1_name ) cmdIndex = 'samtools faidx %s' % ( tmpf1_name ) tmp = tempfile.NamedTemporaryFile( dir=tmpDir ).name tmp_stderr = open( tmp, 'wb' ) proc = subprocess.Popen( args=cmdIndex, shell=True, cwd=tmpDir, 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() #did index succeed? if returncode != 0: raise Exception, 'Error creating index file\n' + stderr cmd = cmd % ( opts, tmpf1_name, tmpf0bam_name, options.output1 ) #perform pileup command tmp = tempfile.NamedTemporaryFile( dir=tmpDir ).name tmp_stderr = open( tmp, 'wb' ) proc = subprocess.Popen( args=cmd, shell=True, cwd=tmpDir, stderr=tmp_stderr.fileno() ) returncode = proc.wait() tmp_stderr.close() #did it succeed? # 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: stop_err( 'Error running Samtools pileup tool\n' + str( e ) ) finally: #clean up temp files if os.path.exists( tmpDir ): shutil.rmtree( tmpDir ) # check that there are results in the output file if os.path.getsize( options.output1 ) > 0: sys.stdout.write( 'Converted BAM to pileup' ) else: stop_err( 'The output file is empty. Your input file may have had no matches, or there may be an error with your input file or settings.' ) if __name__ == "__main__" : __main__()