#!/usr/bin/env python """ Runs BFAST on single-end or paired-end data. TODO: more documentation TODO: - auto-detect gzip or bz2 - split options (?) - queue lengths (?) - assumes reference always has been indexed - main and secondary indexes - scoring matrix file ? - read group file ? usage: bfast_wrapper.py [options] -r, --ref=r: The reference genome to use or index -f, --fastq=f: The fastq file to use for the mapping -F, --output=u: The file to save the output (SAM format) -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) -n, --numThreads=n: The number of threads to use -A, --space=A: The encoding space (0: base 1: color) -o, --offsets=o: The offsets for 'match' -l, --loadAllIndexes=l: Load all indexes into memory -k, --keySize=k: truncate key size in 'match' -K, --maxKeyMatches=K: the maximum number of matches to allow before a key is ignored -M, --maxNumMatches=M: the maximum number of matches to allow before the read is discarded -w, --whichStrand=w: the strands to consider (0: both 1: forward 2: reverse) -t, --timing=t: output timing information to stderr -u, --ungapped=u: performed ungapped local alignment -U, --unconstrained=U: performed local alignment without mask constraints -O, --offset=O: the number of bases before and after each hit to consider in local alignment -q, --avgMismatchQuality=q: average mismatch quality -a, --algorithm=a: post processing algorithm (0: no filtering, 1: all passing filters, 2: unique, 3: best scoring unique, 4: best score all) -P, --disallowPairing=P: do not choose alignments based on pairing -R, --reverse=R: paired end reads are given on reverse strands -z, --random=z: output a random best scoring alignment -D, --dbkey=D: Dbkey for reference genome -H, --suppressHeader=H: Suppress the sam header """ 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 __main__(): parser = optparse.OptionParser() parser.add_option('-r', '--ref', dest='ref', help='The reference genome to index and use') parser.add_option('-f', '--fastq', dest='fastq', help='The fastq file to use for the mapping') parser.add_option('-F', '--output', dest='output', help='The file to save the output (SAM format)') parser.add_option('-A', '--space', dest='space', type="choice", default='0', choices=('0', '1'), help='The encoding space (0: base 1: color)') parser.add_option('-H', '--suppressHeader', action="store_true", dest='suppressHeader', default=False, help='Suppress header') parser.add_option('-n', '--numThreads', dest='numThreads', type="int", default="1", help='The number of threads to use') parser.add_option('-t', '--timing', action="store_true", default=False, dest='timing', help='output timming information to stderr') parser.add_option('-l', '--loadAllIndexes', action="store_true", default=False, dest='loadAllIndexes', help='Load all indexes into memory') parser.add_option('-m', '--indexMask', dest='indexMask', help='String containing info on how to build custom indexes') parser.add_option("-b", "--buildIndex", action="store_true", dest="buildIndex", default=False, help='String containing info on how to build custom indexes') parser.add_option("--indexRepeatMasker", action="store_true", dest="indexRepeatMasker", default=False, help='Do not index lower case sequences. Such as those created by RepeatMasker') parser.add_option('--indexContigOptions', dest='indexContigOptions', default="", help='The contig range options to use for the indexing') parser.add_option('--indexExonsFileName', dest='indexExonsFileName', default="", help='The exons file to use for the indexing') parser.add_option('-o', '--offsets', dest='offsets', default="", help='The offsets for \'match\'') parser.add_option('-k', '--keySize', dest='keySize', type="int", default="-1", help='truncate key size in \'match\'') parser.add_option('-K', '--maxKeyMatches', dest='maxKeyMatches', type="int", default="-1", help='the maximum number of matches to allow before a key is ignored') parser.add_option('-M', '--maxNumMatches', dest='maxNumMatches', type="int", default="-1", help='the maximum number of matches to allow bfore the read is discarded') parser.add_option('-w', '--whichStrand', dest='whichStrand', type="choice", default='0', choices=('0', '1', '2'), help='the strands to consider (0: both 1: forward 2: reverse)') parser.add_option('--scoringMatrixFileName', dest='scoringMatrixFileName', help='Scoring Matrix file used to score the alignments') parser.add_option('-u', '--ungapped', dest='ungapped', action="store_true", default=False, help='performed ungapped local alignment') parser.add_option('-U', '--unconstrained', dest='unconstrained', action="store_true", default=False, help='performed local alignment without mask constraints') parser.add_option('-O', '--offset', dest='offset', type="int", default="0", help='the number of bases before and after each hit to consider in local alignment') parser.add_option('-q', '--avgMismatchQuality', type="int", default="-1", dest='avgMismatchQuality', help='average mismatch quality') parser.add_option('-a', '--algorithm', dest='algorithm', default='0', type="choice", choices=('0', '1', '2', '3', '4'), help='post processing algorithm (0: no filtering, 1: all passing filters, 2: unique, 3: best scoring unique, 4: best score all') parser.add_option('--unpaired', dest='unpaired', action="store_true", default=False, help='do not choose alignments based on pairing') parser.add_option('--reverseStrand', dest='reverseStrand', action="store_true", default=False, help='paired end reads are given on reverse strands') parser.add_option('--pairedEndInfer', dest='pairedEndInfer', action="store_true", default=False, help='break ties when one end of a paired end read by estimating the insert size distribution') parser.add_option('--randomBest', dest='randomBest', action="store_true", default=False, help='output a random best scoring alignment') (options, args) = parser.parse_args() # output version # of tool try: tmp = tempfile.NamedTemporaryFile().name tmp_stdout = open(tmp, 'wb') proc = subprocess.Popen(args='bfast 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('%s\n' % stdout) else: raise Exception except Exception: sys.stdout.write('Could not determine BFAST version\n') buffsize = 1048576 # make temp directory for bfast, requires trailing slash tmp_dir = '%s/' % tempfile.mkdtemp() # 'generic' options used in all bfast commands here if options.timing: all_cmd_options = "-t" else: all_cmd_options = "" try: if options.buildIndex: reference_filepath = tempfile.NamedTemporaryFile(dir=tmp_dir, suffix='.fa').name # build bfast indexes os.symlink(options.ref, reference_filepath) # bfast fast2brg try: nuc_space = ["0"] if options.space == "1": # color space localalign appears to require nuc space brg nuc_space.append("1") for space in nuc_space: cmd = 'bfast fasta2brg -f "%s" -A "%s" %s' % (reference_filepath, space, all_cmd_options) 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 in \'bfast fasta2brg\'.\n' + str(e)) # bfast index try: all_index_cmds = 'bfast index %s -f "%s" -A "%s" -n "%s"' % (all_cmd_options, reference_filepath, options.space, options.numThreads) if options.indexRepeatMasker: all_index_cmds += " -R" if options.indexContigOptions: index_contig_options = [int(_) for _ in options.indexContigOptions.split(',')] if index_contig_options[0] >= 0: all_index_cmds += ' -s "%s"' % index_contig_options[0] if index_contig_options[1] >= 0: all_index_cmds += ' -S "%s"' % index_contig_options[1] if index_contig_options[2] >= 0: all_index_cmds += ' -e "%s"' % index_contig_options[2] if index_contig_options[3] >= 0: all_index_cmds += ' -E "%s"' % index_contig_options[3] elif options.indexExonsFileName: all_index_cmds += ' -x "%s"' % options.indexExonsFileName index_count = 1 for mask, hash_width in [mask.split(':') for mask in options.indexMask.split(',')]: cmd = '%s -m "%s" -w "%s" -i "%i"' % (all_index_cmds, mask, hash_width, index_count) 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) index_count += 1 except Exception as e: raise Exception('Error in \'bfast index\'.\n' + str(e)) else: reference_filepath = options.ref assert reference_filepath and os.path.exists(reference_filepath), 'A valid genome reference was not provided.' # set up aligning and generate aligning command options # set up temp output files tmp_bmf = tempfile.NamedTemporaryFile(dir=tmp_dir) tmp_bmf_name = tmp_bmf.name tmp_bmf.close() tmp_baf = tempfile.NamedTemporaryFile(dir=tmp_dir) tmp_baf_name = tmp_baf.name tmp_baf.close() bfast_match_cmd = 'bfast match -f "%s" -r "%s" -n "%s" -A "%s" -T "%s" -w "%s" %s' % (reference_filepath, options.fastq, options.numThreads, options.space, tmp_dir, options.whichStrand, all_cmd_options) bfast_localalign_cmd = 'bfast localalign -f "%s" -m "%s" -n "%s" -A "%s" -o "%s" %s' % (reference_filepath, tmp_bmf_name, options.numThreads, options.space, options.offset, all_cmd_options) bfast_postprocess_cmd = 'bfast postprocess -O 1 -f "%s" -i "%s" -n "%s" -A "%s" -a "%s" %s' % (reference_filepath, tmp_baf_name, options.numThreads, options.space, options.algorithm, all_cmd_options) if options.offsets: bfast_match_cmd += ' -o "%s"' % options.offsets if options.keySize >= 0: bfast_match_cmd += ' -k "%s"' % options.keySize if options.maxKeyMatches >= 0: bfast_match_cmd += ' -K "%s"' % options.maxKeyMatches if options.maxNumMatches >= 0: bfast_match_cmd += ' -M "%s"' % options.maxNumMatches bfast_localalign_cmd += ' -M "%s"' % options.maxNumMatches if options.scoringMatrixFileName: bfast_localalign_cmd += ' -x "%s"' % options.scoringMatrixFileName bfast_postprocess_cmd += ' -x "%s"' % options.scoringMatrixFileName if options.ungapped: bfast_localalign_cmd += ' -u' if options.unconstrained: bfast_localalign_cmd += ' -U' if options.avgMismatchQuality >= 0: bfast_localalign_cmd += ' -q "%s"' % options.avgMismatchQuality bfast_postprocess_cmd += ' -q "%s"' % options.avgMismatchQuality if options.algorithm == 3: if options.pairedEndInfer: bfast_postprocess_cmd += ' -P' if options.randomBest: bfast_postprocess_cmd += ' -z' if options.unpaired: bfast_postprocess_cmd += ' -U' if options.reverseStrand: bfast_postprocess_cmd += ' -R' # instead of using temp files, should we stream through pipes? bfast_match_cmd += " > %s" % tmp_bmf_name bfast_localalign_cmd += " > %s" % tmp_baf_name bfast_postprocess_cmd += " > %s" % options.output # need to nest try-except in try-finally to handle 2.4 try: # bfast 'match' try: tmp = tempfile.NamedTemporaryFile(dir=tmp_dir).name tmp_stderr = open(tmp, 'wb') proc = subprocess.Popen(args=bfast_match_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 in \'bfast match\'. \n' + str(e)) # bfast 'localalign' try: tmp = tempfile.NamedTemporaryFile(dir=tmp_dir).name tmp_stderr = open(tmp, 'wb') proc = subprocess.Popen(args=bfast_localalign_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 in \'bfast localalign\'. \n' + str(e)) # bfast 'postprocess' try: tmp = tempfile.NamedTemporaryFile(dir=tmp_dir).name tmp_stderr = open(tmp, 'wb') proc = subprocess.Popen(args=bfast_postprocess_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 in \'bfast postprocess\'. \n' + str(e)) # remove header if necessary if options.suppressHeader: 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 as e: raise Exception('Error moving output file before removing headers. \n' + str(e)) fout = open(options.output, 'w') for line in open(tmp_out.name): if len(line) < 3 or line[0:3] not in ['@HD', '@SQ', '@RG', '@PG', '@CO']: fout.write(line) fout.close() # check that there are results in the output file if os.path.getsize(options.output) > 0: if "0" == options.space: sys.stdout.write('BFAST run on Base Space data') else: sys.stdout.write('BFAST run on Color Space data') 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 as e: stop_err('The alignment failed.\n' + str(e)) finally: # clean up temp dir if os.path.exists(tmp_dir): shutil.rmtree(tmp_dir) if __name__ == "__main__": __main__()