def run(self):
"""Subset reads based on read annotation and subset rules."""
infoMsg = "Extracting reads from {f} based on ".format(f=self.inFN)
infoMsg += "rules(FullLength={fl}, nonChimeric={nc}).".format(
fl="true" if self.rules.FL != 0 else "false",
nc="true" if self.rules.nonChimeric != 0 else "false")
logging.info(infoMsg)
if not self.printReadLengthOnly:
with FastaReader(self.inFN) as reader, \
FastaWriter(self.outFN) as writer:
for r in reader:
#print >> sys.stderr, r.name, self.ignore_polyA
annotation = ReadAnnotation.fromString(r.name,
self.ignore_polyA)
if self.satisfy(annotation, self.rules):
writer.writeRecord(r.name, r.sequence)
else: # print read length only, dont print read names and sequences
with FastaReader(self.inFN) as reader, \
open(self.outFN, 'w') as writer:
for r in reader:
annotation = ReadAnnotation.fromString(r.name,
self.ignore_polyA)
if self.satisfy(annotation, self.rules):
writer.write("{rl}\n".format(rl=len(r.sequence)))
def run(self):
"""Subset reads based on read annotation and subset rules."""
infoMsg = "Extracting reads from {f} based on ".format(f=self.inFN)
infoMsg += "rules(FullLength={fl}, nonChimeric={nc}).".format(
fl="true" if self.rules.FL != 0 else "false",
nc="true" if self.rules.nonChimeric != 0 else "false")
logging.info(infoMsg)
if not self.printReadLengthOnly:
with FastaReader(self.inFN) as reader, \
FastaWriter(self.outFN) as writer:
for r in reader:
#print >> sys.stderr, r.name, self.ignore_polyA
annotation = ReadAnnotation.fromString(
r.name, self.ignore_polyA)
if self.satisfy(annotation, self.rules):
writer.writeRecord(r.name, r.sequence)
else: # print read length only, dont print read names and sequences
with FastaReader(self.inFN) as reader, \
open(self.outFN, 'w') as writer:
for r in reader:
annotation = ReadAnnotation.fromString(
r.name, self.ignore_polyA)
if self.satisfy(annotation, self.rules):
writer.write("{rl}\n".format(rl=len(r.sequence)))
def _updateChimeraInfo(self,
suspicous_hits,
in_read_fn,
out_nc_fn,
out_c_fn,
primer_report_fn,
write_report_header=True):
"""
in_read_fn --- a fasta of full-length reads or a fasta of
non-full-length reads.
For each full-length read in in_read_fn FASTA file, detect whether
it is chimeric or not, and write its annotation to
primer_report_fn.
Return:
(num_nc, num_c, num_nc_bases, num_c_bases)
"""
logging.debug(
"Update chimera info for reads in {f} ".format(f=in_read_fn))
logging.debug(
"Write primer report to {rpt}".format(rpt=primer_report_fn))
num_nc, num_c, num_nc_bases, num_c_bases = 0, 0, 0, 0
with FastaReader(in_read_fn) as reader, \
FastaWriter(out_nc_fn) as writer, \
FastaWriter(out_c_fn) as writer_chimera, \
open(primer_report_fn, 'w') as reporter:
if write_report_header:
reporter.write(ReadAnnotation.header(delimiter=",") + "\n")
for r in reader:
# e.g. r.name="movie/zmw/0_100_CCS fiveend=1;threeend=100;"
readid = r.name.split()[0]
annotation = ReadAnnotation.fromString(
r.name, ignore_polyA=self.ignore_polyA)
if readid not in suspicous_hits: # Non-chimeric reads
# Primer of a primer-trimmed read can not be None.
# assert(annotation.primer is not None)
annotation.chimera = 0
num_nc += 1
num_nc_bases += len(r.sequence)
writer.writeRecord(annotation.toAnnotation(), r.sequence)
else: # chimeric reads
annotation.chimera = 1
num_c += 1
num_c_bases += len(r.sequence)
writer_chimera.writeRecord(annotation.toAnnotation(),
r.sequence)
reporter.write(annotation.toReportRecord(delimitor=",") + "\n")
return (num_nc, num_c, num_nc_bases, num_c_bases)
def _updateChimeraInfo(self, suspicous_hits, in_read_fn, out_nc_fn,
out_c_fn, primer_report_fn,
write_report_header=True):
"""
in_read_fn --- a fasta of full-length reads or a fasta of
non-full-length reads.
For each full-length read in in_read_fn FASTA file, detect whether
it is chimeric or not, and write its annotation to
primer_report_fn.
Return:
(num_nc, num_c, num_nc_bases, num_c_bases)
"""
logging.debug("Update chimera info for reads in {f} ".
format(f=in_read_fn))
logging.debug("Write primer report to {rpt}".
format(rpt=primer_report_fn))
num_nc, num_c, num_nc_bases, num_c_bases = 0, 0, 0, 0
with FastaReader(in_read_fn) as reader, \
FastaWriter(out_nc_fn) as writer, \
FastaWriter(out_c_fn) as writer_chimera, \
open(primer_report_fn, 'w') as reporter:
if write_report_header:
reporter.write(ReadAnnotation.header(delimiter=",") + "\n")
for r in reader:
# e.g. r.name="movie/zmw/0_100_CCS fiveend=1;threeend=100;"
readid = r.name.split()[0]
annotation = ReadAnnotation.fromString(r.name,
ignore_polyA=self.ignore_polyA)
if readid not in suspicous_hits: # Non-chimeric reads
# Primer of a primer-trimmed read can not be None.
# assert(annotation.primer is not None)
annotation.chimera = 0
num_nc += 1
num_nc_bases += len(r.sequence)
writer.writeRecord(annotation.toAnnotation(),
r.sequence)
else: # chimeric reads
annotation.chimera = 1
num_c += 1
num_c_bases += len(r.sequence)
writer_chimera.writeRecord(annotation.toAnnotation(),
r.sequence)
reporter.write(annotation.toReportRecord(delimitor=",") + "\n")
return (num_nc, num_c, num_nc_bases, num_c_bases)
def test_satisfy(self):
"""Test function satisfy()."""
inFN = op.join(self.testDir, "data/test_subset.fa")
reads = []
with FastaReader(inFN) as reader:
reads = [x for x in reader]
rules = SubsetRules(1, 1) # Full-length, non-chimeric
obj = ReadsSubsetExtractor("in", "out", rules, True)
ans = [ReadAnnotation.fromString(r.name) for r in reads]
res = [obj.satisfy(an, rules) for an in ans]
expected = [1, 0, 0, 0, 1, 0, 0, 0, 0, 1, 0, 1, 0, 1, 1]
self.assertTrue(res == expected)
def _updateChimeraInfo(self, suspicous_hits, in_read_fn, out_flnc_fn,
out_flc_fn, primer_report_fl_fn):
"""
in_read_fn --- a fasta of full-length reads
For each full-length read in in_read_fn FASTA file, detect whether
it is chimeric or not, and write its annotation to
primer_report_fl_fn.
"""
logging.info("Update chimera info to reads annotations " +
"in the output FASTA file and the primer report.")
with FastaReader(in_read_fn) as reader, \
FastaWriter(out_flnc_fn) as writer, \
FastaWriter(out_flc_fn) as writer_chimera, \
open(primer_report_fl_fn, 'w') as reporter:
reporter.write("\t".join(ReadAnnotation.fieldsNames()) + "\n")
for r in reader:
# e.g. r.name="movie/zmw/0_100_CCS fiveend=1;threeend=100;"
readid = r.name.split()[0]
annotation = ReadAnnotation.fromString(r.name, ignore_polyA=self.ignore_polyA)
if readid not in suspicous_hits: # Non-chimeric reads
# Primer of a primer-trimmed read can not be None.
# assert(annotation.primer is not None)
annotation.chimera = 0
assert(annotation.isFullLength)
self.summary.num_flnc += 1
self.summary.num_flnc_bases += len(r.sequence)
writer.writeRecord(annotation.toAnnotation(),
r.sequence)
else: # chimeric reads
annotation.chimera = 1
self.summary.num_flc += 1
writer_chimera.writeRecord(annotation.toAnnotation(),
r.sequence)
reporter.write(annotation.toReportRecord() + "\n")
def _updateChimeraInfo(self, suspicous_hits, in_read_fn, out_flnc_fn,
out_flc_fn, primer_report_fl_fn):
"""
in_read_fn --- a fasta of full-length reads
For each full-length read in in_read_fn FASTA file, detect whether
it is chimeric or not, and write its annotation to
primer_report_fl_fn.
"""
logging.info("Update chimera info to reads annotations " +
"in the output FASTA file and the primer report.")
with FastaReader(in_read_fn) as reader, \
FastaWriter(out_flnc_fn) as writer, \
FastaWriter(out_flc_fn) as writer_chimera, \
open(primer_report_fl_fn, 'w') as reporter:
reporter.write("\t".join(ReadAnnotation.fieldsNames()) + "\n")
for r in reader:
# e.g. r.name="movie/zmw/0_100_CCS fiveend=1;threeend=100;"
readid = r.name.split()[0]
annotation = ReadAnnotation.fromString(
r.name, ignore_polyA=self.ignore_polyA)
if readid not in suspicous_hits: # Non-chimeric reads
# Primer of a primer-trimmed read can not be None.
# assert(annotation.primer is not None)
annotation.chimera = 0
assert (annotation.isFullLength)
self.summary.num_flnc += 1
self.summary.num_flnc_bases += len(r.sequence)
writer.writeRecord(annotation.toAnnotation(), r.sequence)
else: # chimeric reads
annotation.chimera = 1
self.summary.num_flc += 1
writer_chimera.writeRecord(annotation.toAnnotation(),
r.sequence)
reporter.write(annotation.toReportRecord() + "\n")
def _trimBarCode(self, reads_fn, out_fl_reads_fn, out_nfl_reads_fn,
primer_report_nfl_fn,
best_of_front, best_of_back, primer_indices,
min_seq_len, min_score, change_read_id,
ignore_polyA):
"""Trim bar code from reads in 'reads_fn', annotate each read,
indicating:
whether its 5' primer, 3' primer and polyA tail are seen,
start positions of its 5' primer, 3' primer and polyA tail,
and primer info.
, save non-full-length reads to 'out_nfl_reads_fn',
, save full-length reads to 'out_fl_reads_fn', which can later be
used in chimera detection
, write primer info of nfl reads to _primer_report_nfl_fn.
Note that chimera detection is not necessary for nfl reads, but
is required for fl reads. So we only write primer info for nfl here
and will write primer info for fl reads when chimera detection
is done.
best_of_front/Back: {read_id: {primer_name:DOMRecord}}
min_seq_len: minimum length to output a read.
min_score: minimum score to output a read.
change_read_id: if True, change read ids to 'movie/zmw/start_end'.
"""
logging.info("Trim bar code away from reads.")
logging.debug("Writing full-length trimmed reads to {f}".
format(f=out_fl_reads_fn))
logging.debug("Writing non-full-length trimmed reads to {f}".
format(f=out_nfl_reads_fn))
logging.debug("Writing primer reports before chimera detection to {f}".
format(f=primer_report_nfl_fn))
with FastaReader(reads_fn) as fareader, \
FastaWriter(out_nfl_reads_fn) as nfl_fawriter, \
FastaWriter(out_fl_reads_fn) as fl_fawriter, \
open(primer_report_nfl_fn, 'w') as reporter:
for read in fareader:
self.summary.num_reads += 1 # number of ROI reads
pbread = PBRead(read)
logging.debug("Pick up best primer combo for {r}".
format(r=read.name))
primerIndex, strand, fw, rc = self._pickBestPrimerCombo(
best_of_front[read.name], best_of_back[read.name],
primer_indices, min_score)
logging.debug("read={0}\n".format(read.name) +
"primer={0} strand={1} fw={2} rc={3}".
format(primerIndex, strand, fw, rc))
if fw is None and rc is None:
# No primer seen in this sequence, classified
# as non-full-length
newName = pbread.name
if change_read_id:
newName = "{m}/{z}/{s1}_{e1}{isccs}".format(
m=pbread.movie, z=pbread.zmw,
s1=pbread.start, e1=pbread.end,
isccs=("_CCS" if pbread.isCCS else ""))
annotation = ReadAnnotation(ID=newName)
# Write reports of nfl reads
reporter.write(annotation.toReportRecord(delimitor=",") + "\n")
if len(read.sequence) >= min_seq_len:
# output non-full-length reads to nfl.trimmed.fasta
nfl_fawriter.writeRecord(annotation.toAnnotation(),
read.sequence)
self.summary.num_nfl += 1
else:
self.summary.num_filtered_short_reads += 1
continue
seq = read.sequence if strand == "+" else revcmp(read.sequence)
five_end, three_start = None, None
if fw is not None:
five_end = fw.sEnd
self.summary.num_5_seen += 1
if rc is not None:
three_start = len(seq) - rc.sEnd
self.summary.num_3_seen += 1
s, e = pbread.start, pbread.end
# Try to find polyA tail in read
polyAPos = self._findPolyA(seq, three_start=three_start)
if polyAPos >= 0: # polyA found
seq = seq[:polyAPos]
e1 = s + polyAPos if strand == "+" else e - polyAPos
self.summary.num_polyA_seen += 1
elif three_start is not None: # polyA not found
seq = seq[:three_start]
e1 = s + three_start if strand == "+" else e - three_start
else:
e1 = e if strand == "+" else s
if five_end is not None:
seq = seq[five_end:]
s1 = s + five_end if strand == "+" else e - five_end
else:
s1 = s if strand == "+" else e
newName = pbread.name
if change_read_id:
newName = "{m}/{z}/{s1}_{e1}{isccs}".format(
m=pbread.movie, z=pbread.zmw, s1=s1, e1=e1,
isccs=("_CCS" if pbread.isCCS else ""))
# Create an annotation
annotation = ReadAnnotation(ID=newName, strand=strand,
fiveend=five_end, polyAend=polyAPos,
threeend=three_start, primer=primerIndex,
ignore_polyA=ignore_polyA)
# Write reports for nfl reads
if annotation.isFullLength is not True:
reporter.write(annotation.toReportRecord(delimitor=",") + "\n")
if len(seq) >= min_seq_len:
if annotation.isFullLength is True:
# Write long full-length reads
fl_fawriter.writeRecord(annotation.toAnnotation(), seq)
self.summary.num_fl += 1
else:
# Write long non-full-length reads.
nfl_fawriter.writeRecord(annotation.toAnnotation(), seq)
self.summary.num_nfl += 1
else:
self.summary.num_filtered_short_reads += 1
def _trimBarCode(self, reads_fn, out_fl_reads_fn, out_nfl_reads_fn,
primer_report_nfl_fn,
best_of_front, best_of_back, primer_names,
min_seq_len, min_score, change_read_id,
ignore_polyA, keep_primer):
"""Trim bar code from reads in 'reads_fn', annotate each read,
indicating:
whether its 5' primer, 3' primer and polyA tail are seen,
start positions of its 5' primer, 3' primer and polyA tail,
and primer info.
, save non-full-length reads to 'out_nfl_reads_fn',
, save full-length reads to 'out_fl_reads_fn', which can later be
used in chimera detection
, write primer info of nfl reads to _primer_report_nfl_fn.
Note that chimera detection is not necessary for nfl reads, but
is required for fl reads. So we only write primer info for nfl here
and will write primer info for fl reads when chimera detection
is done.
best_of_front/Back: {read_id: {primer_name:DOMRecord}}
min_seq_len: minimum length to output a read.
min_score: minimum score to output a read.
change_read_id: if True, change read ids to 'movie/zmw/start_end'.
"""
logging.info("Trim bar code away from reads.")
logging.debug("Writing full-length trimmed reads to {f}".
format(f=out_fl_reads_fn))
logging.debug("Writing non-full-length trimmed reads to {f}".
format(f=out_nfl_reads_fn))
logging.debug("Writing primer reports before chimera detection to {f}".
format(f=primer_report_nfl_fn))
with FastaReader(reads_fn) as fareader, \
FastaWriter(out_nfl_reads_fn) as nfl_fawriter, \
FastaWriter(out_fl_reads_fn) as fl_fawriter, \
open(primer_report_nfl_fn, 'w') as reporter:
for read in fareader:
self.summary.num_reads += 1 # number of ROI reads
pbread = PBRead(read)
logging.debug("Pick up best primer combo for {r}".
format(r=read.name))
primerName, strand, fw, rc = self._pickBestPrimerCombo(
best_of_front[read.name], best_of_back[read.name],
primer_names, min_score)
logging.debug("read={0}\n".format(read.name) +
"strand={0} fw={1} rc={2}".
format(strand, fw, rc))
if (strand == '?') or (fw is None and rc is None):
# No primer seen in this sequence, classified
# as non-full-length
newName = pbread.name
if change_read_id:
newName = "{m}/{z}/{s1}_{e1}{isccs}".format(
m=pbread.movie, z=pbread.zmw,
s1=pbread.start, e1=pbread.end,
isccs=("_CCS" if pbread.isCCS else ""))
annotation = ReadAnnotation(ID=newName, primer=primerName)
# Write reports of nfl reads
reporter.write(annotation.toReportRecord(delimitor=",") + "\n")
if len(read.sequence) >= min_seq_len:
# output non-full-length reads to nfl.trimmed.fasta
nfl_fawriter.writeRecord(annotation.toAnnotation(),
read.sequence)
self.summary.num_nfl += 1
else:
self.summary.num_filtered_short_reads += 1
continue
seq = read.sequence if strand == "+" else revcmp(read.sequence)
five_end, three_start = None, None
if fw is not None:
five_end = fw.sEnd
self.summary.num_5_seen += 1
if rc is not None:
three_start = len(seq) - rc.sEnd
self.summary.num_3_seen += 1
s, e = pbread.start, pbread.end
# Try to find polyA tail in read
polyAPos = self._findPolyA(seq, three_start=three_start)
if polyAPos >= 0 and not ignore_polyA: # polyA found and not to ignore it
if not keep_primer:
seq = seq[:polyAPos]
e1 = s + polyAPos if strand == "+" else e - polyAPos
else:
e1 = e if strand == '+' else s
self.summary.num_polyA_seen += 1
elif three_start is not None: # polyA not found but 3' found
if not keep_primer:
seq = seq[:three_start]
e1 = s + three_start if strand == "+" else e - three_start
else:
e1 = e if strand == '+' else s
else: # polyA not found and 3' not found
e1 = e if strand == "+" else s
if five_end is not None:
if not keep_primer:
seq = seq[five_end:]
s1 = s + five_end if strand == "+" else e - five_end
else:
s1 = s if strand == '+' else e
else:
s1 = s if strand == "+" else e
newName = pbread.name
if change_read_id:
newName = "{m}/{z}/{s1}_{e1}{isccs}".format(
m=pbread.movie, z=pbread.zmw, s1=s1, e1=e1,
isccs=("_CCS" if pbread.isCCS else ""))
# Create an annotation
annotation = ReadAnnotation(ID=newName, strand=strand,
fiveend=five_end, polyAend=polyAPos,
threeend=three_start, primer=primerName,
ignore_polyA=ignore_polyA)
# Write reports for nfl reads
if annotation.isFullLength is not True:
reporter.write(annotation.toReportRecord(delimitor=",") + "\n")
if len(seq) >= min_seq_len:
if annotation.isFullLength is True:
# Write long full-length reads
fl_fawriter.writeRecord(annotation.toAnnotation(), seq)
self.summary.num_fl += 1
else:
# Write long non-full-length reads.
nfl_fawriter.writeRecord(annotation.toAnnotation(), seq)
self.summary.num_nfl += 1
else:
self.summary.num_filtered_short_reads += 1