#!/usr/bin/env python """ Runs SRMA on a SAM/BAM file; TODO: more documentation usage: srma_wrapper.py [options] See below for options """ import optparse import os import shutil import subprocess import sys import tempfile def stop_err(msg): sys.stderr.write('%s\n' % msg) sys.exit() def parseRefLoc(refLoc, refUID): for line in open(refLoc): if not line.startswith('#'): fields = line.strip().split('\t') if len(fields) >= 3: if fields[0] == refUID: return fields[1] return None def __main__(): 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('-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() 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 as 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 as 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 as 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: if options.ref: reference_filepath_name = options.ref else: reference_filepath_name = parseRefLoc(options.refLocation, options.refUID) 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: 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) srma_cmds = "%s VALIDATION_STRINGENCY=LENIENT" % srma_cmds # 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 java_opts = '' if '_JAVA_OPTIONS' not in os.environ: java_opts = '-Xmx2048m' cmd = 'java %s -jar %s I=%s O=%s R=%s %s' % (java_opts, 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 as 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 as 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__()