#!/usr/bin/env python """ Runs SRMA on a SAM/BAM file; TODO: more documentation TODO: Allow use of fastq file in user's history, will need to index on-the-fly usage: srma_wrapper.py [options] -r, --ref=r: The reference genome to use or index -i, --input=i: The SAM/BAM input file -o, --output=o: The SAM/BAM output file -O, --offset=O: The alignment offset -Q, --minMappingQuality=Q: The minimum mapping quality -P, --minAlleleProbability=P: The minimum allele probability conditioned on coverage (for the binomial quantile). -C, --minAlleleCoverage=C: The minimum haploid coverage for the consensus. Default value: 3. This option can be set -R, --range=R: A range to examine -c, --correctBases=c: Correct bases -q, --useSequenceQualities=q: Use sequence qualities -M, --maxHeapSize=M: The maximum number of nodes on the heap before re-alignment is ignored -s, --fileSource=s: Whether to use a previously indexed reference sequence or one from history (indexed or history) -p, --params=p: Parameter setting to use (pre_set or full) -D, --dbkey=D: Dbkey for reference genome """ import optparse, os, shutil, subprocess, sys, tempfile def stop_err( msg ): sys.stderr.write( '%s\n' % msg ) sys.exit() def __main__(): #Parse Command Line parser = optparse.OptionParser() parser.add_option( '-r', '--ref', dest='ref', help='The reference genome to index and use' ) parser.add_option( '-u', '--refUID', dest='refUID', help='The pre-index reference genome unique Identifier' ) parser.add_option( '-L', '--refLocations', dest='refLocations', help='The filepath to the srma indices location file' ) parser.add_option( '-i', '--input', dest='input', help='The SAM/BAM input file' ) parser.add_option( '-I', '--inputIndex', dest='inputIndex', help='The SAM/BAM input index file' ) parser.add_option( '-o', '--output', dest='output', help='The SAM/BAM output file' ) parser.add_option( '-O', '--offset', dest='offset', help='The alignment offset' ) parser.add_option( '-Q', '--minMappingQuality', dest='minMappingQuality', help='The minimum mapping quality' ) parser.add_option( '-P', '--minAlleleProbability', dest='minAlleleProbability', help='The minimum allele probability conditioned on coverage (for the binomial quantile).' ) parser.add_option( '-C', '--minAlleleCoverage', dest='minAlleleCoverage', help='The minimum haploid coverage for the consensus' ) parser.add_option( '-R', '--range', dest='range', help='A range to examine' ) parser.add_option( '-c', '--correctBases', dest='correctBases', help='Correct bases ' ) parser.add_option( '-q', '--useSequenceQualities', dest='useSequenceQualities', help='Use sequence qualities ' ) parser.add_option( '-M', '--maxHeapSize', dest='maxHeapSize', help='The maximum number of nodes on the heap before re-alignment is ignored' ) parser.add_option( '-s', '--fileSource', dest='fileSource', help='Whether to use a previously indexed reference sequence or one from history (indexed or history)' ) parser.add_option( '-p', '--params', dest='params', help='Parameter setting to use (pre_set or full)' ) parser.add_option( '-j', '--jarBin', dest='jarBin', default='', help='The path to where jars are stored' ) parser.add_option( '-f', '--jarFile', dest='jarFile', help='The file name of the jar file to use') (options, args) = parser.parse_args() # make temp directory for srma tmp_dir = tempfile.mkdtemp() buffsize = 1048576 # set up reference filenames reference_filepath_name = None # need to create SRMA dict and Samtools fai files for custom genome if options.fileSource == 'history': try: reference_filepath = tempfile.NamedTemporaryFile( dir=tmp_dir, suffix='.fa' ) reference_filepath_name = reference_filepath.name reference_filepath.close() fai_filepath_name = '%s.fai' % reference_filepath_name dict_filepath_name = reference_filepath_name.replace( '.fa', '.dict' ) os.symlink( options.ref, reference_filepath_name ) # create fai file using Samtools index_fai_cmd = 'samtools faidx %s' % reference_filepath_name try: tmp = tempfile.NamedTemporaryFile( dir=tmp_dir ).name tmp_stderr = open( tmp, 'wb' ) proc = subprocess.Popen( args=index_fai_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: # clean up temp dir if os.path.exists( tmp_dir ): shutil.rmtree( tmp_dir ) stop_err( 'Error creating Samtools index for custom genome file: %s\n' % str( e ) ) # create dict file using SRMA dict_cmd = 'java -cp "%s" net.sf.picard.sam.CreateSequenceDictionary R=%s O=%s' % ( os.path.join( options.jarBin, options.jarFile ), reference_filepath_name, dict_filepath_name ) try: tmp = tempfile.NamedTemporaryFile( dir=tmp_dir ).name tmp_stderr = open( tmp, 'wb' ) proc = subprocess.Popen( args=dict_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: # clean up temp dir if os.path.exists( tmp_dir ): shutil.rmtree( tmp_dir ) stop_err( 'Error creating index for custom genome file: %s\n' % str( e ) ) except Exception, e: # clean up temp dir if os.path.exists( tmp_dir ): shutil.rmtree( tmp_dir ) stop_err( 'Problem handling SRMA index (dict file) for custom genome file: %s\n' % str( e ) ) # using built-in dict/index files else: for line in open( options.refLocations ): if not line.startswith( '#' ): fields = line.strip().split( '\t' ) if len( fields ) >= 2: if fields[0] == options.refUID: reference_filepath_name = fields[1] break if reference_filepath_name is None: raise ValueError( 'A valid genome reference was not provided.' ) # set up aligning and generate aligning command options if options.params == 'pre_set': srma_cmds = '' else: if options.useSequenceQualities == 'true': useSequenceQualities = 'true' else: useSequenceQualities = 'false' ranges = 'null' if options.range == 'None': range = 'null' else: range = options.range srma_cmds = "OFFSET=%s MIN_MAPQ=%s MINIMUM_ALLELE_PROBABILITY=%s MINIMUM_ALLELE_COVERAGE=%s RANGES=%s RANGE=%s CORRECT_BASES=%s USE_SEQUENCE_QUALITIES=%s MAX_HEAP_SIZE=%s" % ( options.offset, options.minMappingQuality, options.minAlleleProbability, options.minAlleleCoverage, ranges, range, options.correctBases, options.useSequenceQualities, options.maxHeapSize ) # perform alignments buffsize = 1048576 try: #symlink input bam and index files due to the naming conventions required by srma here input_bam_filename = os.path.join( tmp_dir, '%s.bam' % os.path.split( options.input )[-1] ) os.symlink( options.input, input_bam_filename ) input_bai_filename = "%s.bai" % os.path.splitext( input_bam_filename )[0] os.symlink( options.inputIndex, input_bai_filename ) #create a temp output name, ending in .bam due to required naming conventions? unkown if required output_bam_filename = os.path.join( tmp_dir, "%s.bam" % os.path.split( options.output )[-1] ) # generate commandline cmd = 'java -jar %s I=%s O=%s R=%s %s' % ( os.path.join( options.jarBin, options.jarFile ), input_bam_filename, output_bam_filename, reference_filepath_name, srma_cmds ) # 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' ) 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 executing SRMA. ' + str( e ) # move file from temp location (with .bam name) to provided path shutil.move( output_bam_filename, options.output ) # check that there are results in the output file if os.path.getsize( options.output ) <= 0: 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 re-alignment failed.\n' + str( e ) ) finally: # clean up temp dir if os.path.exists( tmp_dir ): shutil.rmtree( tmp_dir ) if __name__=="__main__": __main__()