def get_strand(seq, probes, thres):
"""Determine which strand the sequence comes from by trying to align probes from the sense strand.
Returns 'sense', 'anti', or None.
Algorithm: This tries each probe in both directions.
If at least one of the alignments has an identity above the threshold, a vote is cast for the
direction with a higher identity.
If the votes that were cast are unanimous for one direction, that strand is returned.
Else, return None."""
votes = []
for probe in probes:
alignment = swalign.smith_waterman(seq, probe)
sense_id = alignment.matches/len(probe)
alignment = swalign.smith_waterman(seq, seqtools.get_revcomp(probe))
anti_id = alignment.matches/len(probe)
# print '{}: sense: {}, anti: {}'.format(probe, sense_id, anti_id)
if sense_id > thres or anti_id > thres:
if sense_id > anti_id:
votes.append('sense')
else:
votes.append('anti')
strand = None
for vote in votes:
if strand:
if strand != vote:
return None
else:
strand = vote
return strand
def get_strand(seq, probes, thres):
"""Determine which strand the sequence comes from by trying to align probes from the sense strand.
Returns 'sense', 'anti', or None.
Algorithm: This tries each probe in both directions.
If at least one of the alignments has an identity above the threshold, a vote is cast for the
direction with a higher identity.
If the votes that were cast are unanimous for one direction, that strand is returned.
Else, return None."""
votes = []
for probe in probes:
alignment = swalign.smith_waterman(seq, probe)
sense_id = alignment.matches/len(probe)
alignment = swalign.smith_waterman(seq, seqtools.get_revcomp(probe))
anti_id = alignment.matches/len(probe)
# print '{}: sense: {}, anti: {}'.format(probe, sense_id, anti_id)
if sense_id > thres or anti_id > thres:
if sense_id > anti_id:
votes.append('sense')
else:
votes.append('anti')
strand = None
for vote in votes:
if strand:
if strand != vote:
return None
else:
strand = vote
return strand
def is_alignment_reversed(barcode1, barcode2):
"""Return True if the barcodes are reversed with respect to each other, False otherwise.
"reversed" in this case meaning the alpha + beta halves are swapped.
Determine by aligning the two to each other, once in their original forms, and once with the
second barcode reversed. If the smith-waterman score is higher in the reversed form, return True.
"""
half = len(barcode2)//2
barcode2_rev = barcode2[half:] + barcode2[:half]
fwd_align = swalign.smith_waterman(barcode1, barcode2)
rev_align = swalign.smith_waterman(barcode1, barcode2_rev)
if rev_align.score > fwd_align.score:
return True
else:
return False
def make_dcss(sscss):
# ordermates is the mapping between the duplex consensus mate number and the order/mates of the
# SSCSs it's composed of. It's arbitrary but consistent, to make sure the duplex consensuses have
# different mate numbers, and they're the same from run to run.
ordermates = {
0: (('ab', 0), ('ba', 1)),
1: (('ab', 1), ('ba', 0)),
}
# Get the consensus of each pair of SSCSs.
# dcss is indexed by (0-based) mate.
dcss = []
for duplex_mate in 0, 1:
# Gather the pair of reads for this duplex consensus.
sscs_pair = []
for order, mate in ordermates[duplex_mate]:
sscs = sscss.get((order, mate))
if sscs:
sscs_pair.append(sscs)
if len(sscs_pair) < 2:
# If we didn't find two SSCSs for this duplex mate, we can't make a complete pair of duplex
# consensus sequences.
break
align = swalign.smith_waterman(sscs_pair[0]['seq'],
sscs_pair[1]['seq'])
if len(align.target) != len(align.query):
message = '{} != {}:\n'.format(len(align.target), len(align.query))
message += '\n'.join([repr(sscs) for sscs in sscs_pair])
raise AssertionError(message)
seq = consensus.build_consensus_duplex_simple(align.target,
align.query)
reads_per_strand = [sscs['nreads'] for sscs in sscs_pair]
dcss.append({'seq': seq, 'nreads': reads_per_strand})
assert len(dcss) == 0 or len(dcss) == 2, len(dcss)
return dcss
def make_dcss(sscss):
# ordermates is the mapping between the duplex consensus mate number and the order/mates of the
# SSCSs it's composed of. It's arbitrary but consistent, to make sure the duplex consensuses have
# different mate numbers, and they're the same from run to run.
ordermates = {
0: (('ab', 0), ('ba', 1)),
1: (('ab', 1), ('ba', 0)),
}
# Get the consensus of each pair of SSCSs.
# dcss is indexed by (0-based) mate.
dcss = []
for duplex_mate in 0, 1:
# Gather the pair of reads for this duplex consensus.
sscs_pair = []
for order, mate in ordermates[duplex_mate]:
sscs = sscss.get((order, mate))
if sscs:
sscs_pair.append(sscs)
if len(sscs_pair) < 2:
# If we didn't find two SSCSs for this duplex mate, we can't make a complete pair of duplex
# consensus sequences.
break
align = swalign.smith_waterman(sscs_pair[0]['seq'], sscs_pair[1]['seq'])
if len(align.target) != len(align.query):
message = '{} != {}:\n'.format(len(align.target), len(align.query))
message += '\n'.join([repr(sscs) for sscs in sscs_pair])
raise AssertionError(message)
seq = consensus.build_consensus_duplex_simple(align.target, align.query)
reads_per_strand = [sscs['nreads'] for sscs in sscs_pair]
dcss.append({'seq':seq, 'nreads':reads_per_strand})
assert len(dcss) == 0 or len(dcss) == 2, len(dcss)
return dcss
def get_duplex_consensi(family):
consensi = []
for (order1, mate1), (order2, mate2) in (('ab', 0), ('ba', 1)), (('ab', 1), ('ba', 0)):
result = swalign.smith_waterman(family[order1][mate1].consensus.replace('-', ''),
family[order2][mate2].consensus.replace('-', ''))
consensi.append(consensuslib.build_consensus_duplex_simple(result.query, result.target))
return consensi
def process_duplex(duplex, barcode, workers=None, stats=None, incl_sscs=False, sscs_fh=None,
processes=1, min_reads=1, qual_thres=' '):
stats['families'] += 1
# Are we the controller process or a worker?
if processes > 1:
i = stats['families'] % len(workers)
worker = workers[i]
delegate(worker, duplex, barcode)
return
# We're a worker. Actually process the family.
start = time.time()
consensi = []
reads_per_strand = []
duplex_mate = None
for (order, mate), family in duplex.items():
reads = len(family)
if reads < min_reads:
continue
# The mate number for the duplex consensus. It's arbitrary, but all that matters is that the
# two mates have different numbers. This system ensures that:
# Mate 1 is from the consensus of ab/1 and ba/2 families, while mate 2 is from ba/1 and ab/2.
if (order == 'ab' and mate == 1) or (order == 'ba' and mate == 2):
duplex_mate = 1
else:
duplex_mate = 2
seqs = [read['seq'] for read in family]
quals = [read['qual'] for read in family]
consensi.append(consensus.get_consensus(seqs, quals, qual_thres=qual_thres))
reads_per_strand.append(reads)
assert len(consensi) <= 2
if sscs_fh:
for cons, (order, mate), reads in zip(consensi, duplex.keys(), reads_per_strand):
sscs_fh.write('>{bar}.{order}.{mate} {reads}\n'.format(bar=barcode, order=order, mate=mate,
reads=reads))
sscs_fh.write(cons+'\n')
if len(consensi) == 1 and incl_sscs:
print_duplex(consensi[0], barcode, duplex_mate, reads_per_strand)
elif len(consensi) == 2:
align = swalign.smith_waterman(*consensi)
#TODO: log error & return if len(align.target) != len(align.query)
cons = consensus.build_consensus_duplex_simple(align.target, align.query)
print_duplex(cons, barcode, duplex_mate, reads_per_strand)
elapsed = time.time() - start
logging.info('{} sec for {} reads.'.format(elapsed, sum(reads_per_strand)))
if stats and len(consensi) > 0:
stats['time'] += elapsed
stats['reads'] += sum(reads_per_strand)
stats['runs'] += 1
def get_similarity(seq1, seq2):
align = swalign.smith_waterman(seq1, seq2)
logging.debug(align.target + '\n' + align.query)
return align.matches / len(align.query)
def get_similarity(seq1, seq2): align = swalign.smith_waterman(seq1, seq2) logging.debug(align.target+'\n'+align.query) return align.matches / len(align.query)
