def readAndIndex(iterator, with_value=True):
'''read from gtf stream and index.
returns an :class:`IndexedGenome.IndexedGenome`
'''
if with_value:
index = IndexedGenome.IndexedGenome()
for gtf in iterator:
index.add(gtf.contig, gtf.start, gtf.end, gtf)
else:
index = IndexedGenome.Simple()
for gtf in iterator:
index.add(gtf.contig, gtf.start, gtf.end)
return index
def readIntervals(infile, options):
ninput = 0
t = time.time()
if options.format == "gtf":
index = IndexedGenome.IndexedGenome()
for gffs in GTF.transcript_iterator(GTF.iterator(infile)):
ali = alignlib_lite.py_makeAlignmentBlocks()
for gff in gffs:
if gff.feature != "exon":
continue
ali.addDiagonal(gff.start, gff.end, 0)
index.add(min([x.start for x in gffs]),
max([x.end for x in gffs]),
ali)
ninput += 1
if ninput % options.report_step == 0:
E.info(
"reading intervals - progress: ninput=%i, time=%i, avg=%f"
% (ninput,
time.time() - t, float(time.time() - t) / ninput))
elif options.format == "gff":
index = IndexedGenome.Simple()
for g in GTF.iterator(infile):
index.add(g.contig, g.start, g.end)
ninput += 1
if ninput % options.report_step == 0:
E.info(
"reading intervals - progress: ninput=%i, time=%i, avg=%f"
% (ninput, time.time() - t,
float(time.time() - t) / ninput))
E.info("read intervals: %i contigs, %i intervals" % (len(index), ninput))
return index
def main(argv=None):
"""script main.
parses command line options in sys.argv, unless *argv* is given.
"""
if not argv:
argv = sys.argv
# setup command line parser
parser = E.OptionParser(version="%prog version: $Id$",
usage=globals()["__doc__"])
parser.add_option(
"-g", "--gtf-file", dest="filename_gtf", type="string",
help="filename with gene models in gtf format [%default]")
parser.add_option(
"-m", "--filename-mismapped", dest="filename_mismapped", type="string",
help="output bam file for mismapped reads [%default]")
parser.add_option(
"-j", "--junctions-bed-file", dest="filename_junctions", type="string",
help="bam file with reads mapped across junctions [%default]")
parser.add_option(
"-r", "--filename-regions", dest="filename_regions", type="string",
help="filename with regions to remove in bed format [%default]")
parser.add_option(
"-t", "--transcripts-gtf-file", dest="filename_transcriptome",
type="string",
help="bam file with reads mapped against transcripts [%default]")
parser.add_option(
"-p", "--map-tsv-file", dest="filename_map", type="string",
help="filename mapping transcript numbers (used by "
"--filename-transciptome) to transcript names "
"(used by --filename-gtf) [%default]")
parser.add_option(
"-s", "--filename-stats", dest="filename_stats", type="string",
help="filename to output stats to [%default]")
parser.add_option(
"-o", "--colour",
dest="colour_mismatches", action="store_true",
help="mismatches will use colour differences (CM tag) [%default]")
parser.add_option(
"-i", "--ignore-mismatches",
dest="ignore_mismatches", action="store_true",
help="ignore mismatches [%default]")
parser.add_option(
"-c", "--remove-contigs", dest="remove_contigs", type="string",
help="','-separated list of contigs to remove [%default]")
parser.add_option(
"-f", "--force-output", dest="force", action="store_true",
help="force overwriting of existing files [%default]")
parser.add_option("-u", "--unique", dest="unique", action="store_true",
help="remove reads not matching uniquely [%default]")
parser.add_option("--output-sam", dest="output_sam", action="store_true",
help="output in sam format [%default]")
parser.set_defaults(
filename_gtf=None,
filename_mismapped=None,
filename_junctions=None,
filename_transcriptome=None,
filename_map=None,
remove_contigs=None,
force=False,
unique=False,
colour_mismatches=False,
ignore_mismatches=False,
output_sam=False,
filename_table=None,
)
# add common options (-h/--help, ...) and parse command line
(options, args) = E.start(parser, argv=argv)
if len(args) != 1:
raise ValueError("please supply one bam file")
bamfile_genome = args[0]
genome_samfile = pysam.AlignmentFile(bamfile_genome, "rb")
if options.remove_contigs:
options.remove_contigs = options.remove_contigs.split(",")
if options.filename_map:
E.info("reading map")
id_map = IOTools.read_map(
IOTools.open_file(options.filename_map), has_header=True)
id_map = dict([(y, x) for x, y in id_map.items()])
else:
id_map = None
transcripts = {}
if options.filename_gtf:
E.info("indexing geneset")
mapped, missed = 0, 0
for gtf in GTF.transcript_iterator(
GTF.iterator(IOTools.open_file(options.filename_gtf))):
gtf.sort(key=lambda x: x.start)
transcript_id = gtf[0].transcript_id
if id_map:
try:
transcript_id = id_map[transcript_id]
mapped += 1
except KeyError:
missed += 1
continue
transcripts[transcript_id] = gtf
E.info("read %i transcripts from geneset (%i mapped, %i missed)" %
(len(transcripts), mapped, missed))
regions_to_remove = None
if options.filename_regions:
E.info("indexing regions")
regions_to_remove = IndexedGenome.Simple()
for bed in Bed.iterator(IOTools.open_file(options.filename_regions)):
regions_to_remove.add(bed.contig, bed.start, bed.end)
E.info("read %i regions" % len(regions_to_remove))
if options.filename_transcriptome:
transcripts_samfile = pysam.AlignmentFile(options.filename_transcriptome,
"rb")
else:
transcripts_samfile = None
if options.output_sam:
output_samfile = pysam.AlignmentFile("-", "wh", template=genome_samfile)
else:
output_samfile = pysam.AlignmentFile("-", "wb", template=genome_samfile)
if options.filename_mismapped:
if not options.force and os.path.exists(options.filename_mismapped):
raise IOError("output file %s already exists" %
options.filename_mismapped)
output_mismapped = pysam.AlignmentFile(options.filename_mismapped,
"wb",
template=genome_samfile)
else:
output_mismapped = None
if options.filename_junctions:
junctions_samfile = pysam.AlignmentFile(options.filename_junctions,
"rb")
else:
junctions_samfile = None
c = bams2bam_filter(genome_samfile,
output_samfile,
output_mismapped,
transcripts_samfile,
junctions_samfile,
transcripts,
regions=regions_to_remove,
unique=options.unique,
remove_contigs=options.remove_contigs,
colour_mismatches=options.colour_mismatches,
ignore_mismatches=options.ignore_mismatches,
ignore_transcripts=transcripts_samfile is None,
ignore_junctions=junctions_samfile is None)
if options.filename_stats:
outf = IOTools.open_file(options.filename_stats, "w")
outf.write("category\tcounts\n%s\n" % c.asTable())
outf.close()
if options.filename_transcriptome:
transcripts_samfile.close()
genome_samfile.close()
output_samfile.close()
if output_mismapped:
output_mismapped.close()
# write footer and output benchmark information.
E.stop()
def main(argv=None):
"""script main.
parses command line options in sys.argv, unless *argv* is given.
"""
if not argv:
argv = sys.argv
# setup command line parser
parser = E.OptionParser(version="%prog version: $Id",
usage=globals()["__doc__"])
parser.add_option("--bed-file",
dest="infiles",
type="string",
metavar="bed",
help="supply list of bed files",
action="append")
parser.set_defaults(infiles=[])
# add common options (-h/--help, ...) and parse command line
(options, args) = E.start(parser, argv=argv)
options.infiles.extend(args)
if len(options.infiles) == 0:
raise ValueError('please provide at least 1 bed file')
E.info("concatenating bed files")
# concatenate the list of files
tmp = tempfile.NamedTemporaryFile(delete=False, mode="w")
tmp_merge = tempfile.NamedTemporaryFile(delete=False, mode="w")
infs = options.infiles
for inf in infs:
for bed in Bed.iterator(IOTools.open_file(inf)):
tmp.write("%s\n" % bed)
tmp.close()
E.info("merging bed entries")
# merge the bed entries in the file
name = tmp.name
tmp_bed = pybedtools.BedTool(name)
tmp_bed.sort().merge().saveas(tmp_merge.name)
tmp_merge.close()
E.info("indexing bed entries")
# index the bed entries
merged = IndexedGenome.Simple()
for bed in Bed.iterator(IOTools.open_file(tmp_merge.name)):
merged.add(bed.contig, bed.start, bed.end)
counts = collections.defaultdict(int)
# list of samples
samples = options.infiles
E.info("counting no. samples overlapping each interval")
for sample in samples:
found = set()
for bed in Bed.iterator(IOTools.open_file(sample)):
if merged.contains(bed.contig, bed.start, bed.end):
key = [bed.contig] + \
[x for x in merged.get(bed.contig, bed.start, bed.end)]
key = (key[0], key[1][0], key[1][1])
if key in found:
continue
found.add(key)
# tuple of interval description as key - (contig, start, end)
counts[key] += 1
# open outfile
options.stdout.write("contig\tstart\tend\tcount\n")
E.info("outputting result")
for interval, count in sorted(counts.items()):
options.stdout.write("\t".join(map(str, interval)) + "\t" +
str(count) + "\n")
# write footer and output benchmark information.
E.stop()
