def resolved_tool_contract_runner(resolved_contract):
rc = resolved_contract
alignment_path = rc.task.input_files[0]
reference_path = rc.task.input_files[1]
gff_path = rc.task.output_files[0]
vcf_path = rc.task.output_files[1]
dataset_path = rc.task.output_files[2]
fasta_path = re.sub(".contigset.xml", ".fasta", dataset_path)
fastq_path = rc.task.output_files[3]
args = [
alignment_path,
"--verbose",
"--reference", reference_path,
"--outputFilename", gff_path,
"--outputFilename", fasta_path,
"--outputFilename", fastq_path,
"--outputFilename", vcf_path,
"--numWorkers", str(rc.task.nproc),
"--minCoverage", str(rc.task.options[Constants.MIN_COVERAGE_ID]),
"--minConfidence", str(rc.task.options[Constants.MIN_CONFIDENCE_ID]),
"--maskRadius", str(Constants.DEFAULT_MASK_RADIUS) if \
bool(rc.task.options[Constants.MASKING_ID]) else "0",
"--algorithm", rc.task.options[Constants.ALGORITHM_ID],
"--alignmentSetRefWindows",
]
args_ = get_parser().arg_parser.parser.parse_args(args)
rc = args_runner(args_)
if rc == 0:
pysam.faidx(fasta_path)
ds = ContigSet(fasta_path, strict=True)
ds.write(dataset_path)
return rc
def run(referenceset, fastq, gff, fasta, contigset, alignmentset, options,
log_level):
#'--log-file foo.log',
#'--verbose',
#'--debug', # requires 'ipdb'
#'-j NWORKERS',
#'--algorithm quiver',
#'--diploid', # binary
#'--minConfidence 40',
#'--minCoverage 5',
#'--alignmentSetRefWindows',
cmd = "variantCaller --log-level {log_level} {options} --referenceFilename {referenceset} -o {fastq} -o {gff} -o {fasta} {alignmentset}"
system(cmd.format(**locals()))
try:
say('Converting fasta {!r} to contigset {!r}'.format(fasta, contigset))
# Convert to contigset.xml
import pysam
pysam.faidx(fasta) # pylint: disable=no-member
# I do not know why pylint does not see this defined.
ds = ContigSet(fasta, strict=True)
ds.write(contigset, relPaths=True)
say('Successfully wrapped fasta {!r} in contigset {!r}'.format(
fasta, contigset))
except Exception:
say(traceback.format_exc())
say('Skipping conversion to contigset.')
def test_contigset_build(self):
ds1 = ContigSet(data.getXml(3), skipMissing=True)
assert type(ds1).__name__ == 'ContigSet'
assert type(ds1._metadata).__name__ == 'ContigSetMetadata'
ds2 = ContigSet(data.getXml(3), skipMissing=True)
assert type(ds2).__name__ == 'ContigSet'
assert type(ds2._metadata).__name__ == 'ContigSetMetadata'
def resolved_tool_contract_runner(resolved_contract):
rc = resolved_contract
alignment_path = rc.task.input_files[0]
reference_path = rc.task.input_files[1]
gff_path = rc.task.output_files[0]
vcf_path = rc.task.output_files[1]
dataset_path = rc.task.output_files[2]
fasta_path = re.sub(".contigset.xml", ".fasta", dataset_path)
fastq_path = rc.task.output_files[3]
args = [
alignment_path,
"--verbose",
"--reference", reference_path,
"--outputFilename", gff_path,
"--outputFilename", fasta_path,
"--outputFilename", fastq_path,
"--outputFilename", vcf_path,
"--numWorkers", str(rc.task.nproc),
"--minCoverage", str(rc.task.options[Constants.MIN_COVERAGE_ID]),
"--minConfidence", str(rc.task.options[Constants.MIN_CONFIDENCE_ID]),
"--maskRadius", str(Constants.DEFAULT_MASK_RADIUS) if \
bool(rc.task.options[Constants.MASKING_ID]) else "0",
"--algorithm", rc.task.options[Constants.ALGORITHM_ID],
"--alignmentSetRefWindows",
]
args_ = get_parser().arg_parser.parser.parse_args(args)
rc = args_runner(args_)
if rc == 0:
pysam.faidx(fasta_path)
ds = ContigSet(fasta_path, strict=True)
ds.write(dataset_path)
return rc
def run(referenceset, fastq, gff, fasta, contigset, alignmentset, options, log_level):
#'--log-file foo.log',
#'--verbose',
#'--debug', # requires 'ipdb'
#'-j NWORKERS',
#'--algorithm quiver',
#'--diploid', # binary
#'--minConfidence 40',
#'--minCoverage 5',
#'--alignmentSetRefWindows',
cmd = "variantCaller --log-level {log_level} {options} --referenceFilename {referenceset} -o {fastq} -o {gff} -o {fasta} {alignmentset}"
system(cmd.format(**locals()))
try:
say('Converting fasta {!r} to contigset {!r}'.format(fasta, contigset))
# Convert to contigset.xml
import pysam
pysam.faidx(fasta) # pylint: disable=no-member
# I do not know why pylint does not see this defined.
ds = ContigSet(fasta, strict=True)
ds.write(contigset, relPaths=True)
say('Successfully wrapped fasta {!r} in contigset {!r}'.format(fasta, contigset))
except Exception:
say(traceback.format_exc())
say('Skipping conversion to contigset.')
def _get_fasta_path(file_name):
if file_name.endswith(".contigset.xml"):
ds = ContigSet(file_name)
fasta_files = ds.toExternalFiles()
assert len(fasta_files) == 1
return fasta_files[0]
return file_name
def test_contigset_build(self):
ds1 = ContigSet(data.getXml(3), skipMissing=True)
self.assertEquals(type(ds1).__name__, 'ContigSet')
self.assertEquals(type(ds1._metadata).__name__, 'ContigSetMetadata')
ds2 = ContigSet(data.getXml(3), skipMissing=True)
self.assertEquals(type(ds2).__name__, 'ContigSet')
self.assertEquals(type(ds2._metadata).__name__, 'ContigSetMetadata')
def test_contigset_consolidate_int_names(self):
# build set to merge
outdir = tempfile.mkdtemp(suffix="dataset-unittest")
inFas = os.path.join(outdir, 'infile.fasta')
outFas1 = os.path.join(outdir, 'tempfile1.fasta')
outFas2 = os.path.join(outdir, 'tempfile2.fasta')
# copy fasta reference to hide fai and ensure FastaReader is used
shutil.copyfile(
ReferenceSet(data.getXml(8)).toExternalFiles()[0], inFas)
rs1 = ContigSet(inFas)
double = 'B.cereus.1'
exp_double = rs1.get_contig(double)
# todo: modify the names first:
with FastaWriter(outFas1) as writer:
writer.writeRecord('5141', exp_double.sequence)
with FastaWriter(outFas2) as writer:
writer.writeRecord('5142', exp_double.sequence)
exp_double_seqs = [exp_double.sequence, exp_double.sequence]
exp_names = ['5141', '5142']
obs_file = ContigSet(outFas1, outFas2)
log.debug(obs_file.toExternalFiles())
obs_file.consolidate()
log.debug(obs_file.toExternalFiles())
# open obs and compare to exp
for name, seq in zip(exp_names, exp_double_seqs):
assert obs_file.get_contig(name).sequence[:] == seq
def resolved_tool_contract_runner(resolved_contract):
rc = resolved_contract
alignment_path = rc.task.input_files[0]
reference_path = rc.task.input_files[1]
gff_path = rc.task.output_files[0]
dataset_path = rc.task.output_files[1]
fasta_path = re.sub(".contigset.xml", ".fasta", dataset_path)
fastq_path = rc.task.output_files[2]
args = [
alignment_path,
"--verbose",
"--reference", reference_path,
"--outputFilename", gff_path,
"--outputFilename", fasta_path,
"--outputFilename", fastq_path,
"--numWorkers", str(rc.task.nproc),
"--minCoverage", str(rc.task.options[Constants.MIN_COVERAGE_ID]),
"--minConfidence", str(rc.task.options[Constants.MIN_CONFIDENCE_ID]),
"--algorithm", rc.task.options[Constants.ALGORITHM_ID],
"--alignmentSetRefWindows",
]
if rc.task.options[Constants.DIPLOID_MODE_ID]:
args.append("--diploid")
args_ = get_parser().arg_parser.parser.parse_args(args)
rc = args_runner(args_)
if rc == 0:
pysam.faidx(fasta_path)
ds = ContigSet(fasta_path, strict=True)
ds.write(dataset_path)
return rc
def _get_fasta_path(file_name):
if file_name.endswith(".contigset.xml"):
ds = ContigSet(file_name)
fasta_files = ds.toExternalFiles()
assert len(fasta_files) == 1
return fasta_files[0]
return file_name
def test_contigset_consolidate_genomic_consensus(self):
"""
Verify that the contigs output by GenomicConsensus (e.g. quiver) can
be consolidated.
"""
FASTA1 = ("lambda_NEB3011_0_60",
"GGGCGGCGACCTCGCGGGTTTTCGCTATTTATGAAAATTTTCCGGTTTAAGGCGTTTCCG")
FASTA2 = ("lambda_NEB3011_120_180",
"CACTGAATCATGGCTTTATGACGTAACATCCGTTTGGGATGCGACTGCCACGGCCCCGTG")
FASTA3 = ("lambda_NEB3011_60_120",
"GTGGACTCGGAGCAGTTCGGCAGCCAGCAGGTGAGCCGTAATTATCATCTGCGCGGGCGT")
files = []
for i, (header, seq) in enumerate([FASTA1, FASTA2, FASTA3]):
_files = []
for suffix in ["", "|quiver", "|plurality", "|arrow", "|poa"]:
tmpfile = tempfile.NamedTemporaryFile(suffix=".fasta").name
with open(tmpfile, "w") as f:
f.write(">{h}{s}\n{q}".format(h=header, s=suffix, q=seq))
_files.append(tmpfile)
files.append(_files)
for i in range(3):
ds = ContigSet(*[f[i] for f in files])
out1 = tempfile.NamedTemporaryFile(suffix=".contigset.xml").name
fa1 = tempfile.NamedTemporaryFile(suffix=".fasta").name
ds.consolidate(fa1)
ds.write(out1)
with ContigSet(out1) as ds_new:
self.assertEqual(len([rec for rec in ds_new]), 1,
"failed on %d" % i)
def _write_fasta_or_contigset(file_name):
fasta_file = re.sub(".contigset.xml", ".fasta", file_name)
rec = [">chr%d\nacgtacgtacgt" % x for x in range(251)]
with open(fasta_file, "w") as f:
f.write("\n".join(rec))
if file_name.endswith(".xml"):
cs = ContigSet(fasta_file)
cs.write(file_name)
def _write_fasta_or_contigset(file_name):
fasta_file = re.sub(".contigset.xml", ".fasta", file_name)
rec = [">chr%d\nacgtacgtacgt" % x for x in range(251)]
with open(fasta_file, "w") as f:
f.write("\n".join(rec))
if file_name.endswith(".xml"):
cs = ContigSet(fasta_file)
cs.write(file_name)
def test_contigset_build(self):
ds1 = ContigSet(data.getXml(3))
self.assertEquals(type(ds1).__name__, 'ContigSet')
self.assertEquals(type(ds1._metadata).__name__, 'ContigSetMetadata')
ds2 = ContigSet(data.getXml(3))
self.assertEquals(type(ds2).__name__, 'ContigSet')
self.assertEquals(type(ds2._metadata).__name__, 'ContigSetMetadata')
for contigmd in ds2.metadata.contigs:
self.assertEquals(type(contigmd).__name__, 'ContigMetadata')
def makeReport(inReadsFN, inSummaryFN, outDir):
"""
Generate a report with ID, tables, attributes and plot groups.
inReadsFN --- an input FASTA file which has all consensus
isoforms produced by pbtranscript.py cluster.
This file is required to plot a read length histogram as part of
the report:
consensus_isoforms_readlength_hist.png
inSummaryFN --- a summary TXT file with cluster attributes,
including two attributes:
number of consensus isoforms
average length of consensus isoforms
Attributes of the report are extracted from this file.
"""
log.info("Plotting read length histogram from file: {f}".
format(f=inReadsFN))
# Collect read lengths of
reader = ContigSet(inReadsFN)
rs = [len(r.sequence) for r in reader]
reader.close()
readlengths = np.array(rs)
# Plot read length histogram
readlength_plot = create_readlength_plot(readlengths, outDir)
readlength_group = PlotGroup(Constants.PG_READLENGTH,
title="Read Length of Consensus Isoforms Reads",
plots=[readlength_plot],
thumbnail=readlength_plot.thumbnail)
log.info("Plotting summary attributes from file: {f}".
format(f=inSummaryFN))
# Produce attributes based on summary.
dataset_uuids = [ContigSet(inReadsFN).uuid]
if inSummaryFN.endswith(".json"):
attributes = _report_to_attributes(inSummaryFN)
r = load_report_from_json(inSummaryFN)
# FIXME(nechols)(2016-03-22) not using the dataset UUIDs from these
# reports; should we be?
else:
attributes = summaryToAttributes(inSummaryFN)
table = attributesToTable(attributes)
log.info(str(table))
# A report is consist of ID, tables, attributes, and plotgroups.
report = Report(Constants.R_ID,
title="Transcript Clustering",
attributes=attributes,
plotgroups=[readlength_group],
dataset_uuids=dataset_uuids)
return report
def __gather_contigset(resource_file_extension, input_files, output_file,
new_resource_file=None,
skip_empty=True):
"""
:param input_files: List of file paths
:param output_file: File Path
:param new_resource_file: the path of the file to which the other contig
files are consolidated
:param skip_empty: Ignore empty files (doesn't do much yet)
:return: Output file
:rtype: str
"""
if skip_empty:
_input_files = []
for file_name in input_files:
cs = ContigSet(file_name)
if len(cs.toExternalFiles()) > 0:
_input_files.append(file_name)
input_files = _input_files
tbr = ContigSet(*input_files)
if not new_resource_file:
if output_file.endswith('xml'):
new_resource_file = output_file[:-3] + resource_file_extension
tbr.consolidate(new_resource_file)
tbr.newUuid()
tbr.write(output_file)
return output_file
def gather_contigset(input_files,
output_file,
new_resource_file=None,
skip_empty=True):
"""
:param input_files: List of file paths
:param output_file: File Path
:param new_resource_file: the path of the file to which the other contig
files are consolidated
:param skip_empty: Ignore empty files (doesn't do much yet)
:return: Output file
:rtype: str
"""
if skip_empty:
_input_files = []
for file_name in input_files:
cs = ContigSet(file_name)
if len(cs.toExternalFiles()) > 0:
_input_files.append(file_name)
input_files = _input_files
tbr = ContigSet(*input_files)
if not new_resource_file:
if output_file.endswith('xml'):
new_resource_file = output_file[:-3] + 'fasta'
tbr.consolidate(new_resource_file)
tbr.write(output_file)
return output_file
def _write_fasta_or_contigset(file_name, make_faidx=False, n_records=251):
fasta_file = re.sub(".contigset.xml", ".fasta", file_name)
rec = [">chr%d\nacgtacgtacgt" % x for x in range(n_records)]
with open(fasta_file, "w") as f:
f.write("\n".join(rec))
f.flush()
if make_faidx:
pysam.faidx(fasta_file)
if file_name.endswith(".xml"):
cs = ContigSet(fasta_file, strict=make_faidx)
cs.write(file_name)
def write_contigset_records(pbcore_writer_class, records, file_name):
"""
Writes the Chunked fasta files and Writes a ContigSet.xml
Filename has contigset.xml
"""
fasta_file_name = ".".join(file_name.split(".")[:-2]) + ".fasta"
write_pbcore_records(pbcore_writer_class, records, fasta_file_name)
log.debug("Writing ContigSet XML to {f}".format(f=file_name))
ds = ContigSet(fasta_file_name)
ds.write(file_name)
def write_contigset_records(pbcore_writer_class, records, file_name):
"""
Writes the Chunked fasta files and Writes a ContigSet.xml
Filename has contigset.xml
"""
fasta_file_name = ".".join(file_name.split(".")[:-2]) + ".fasta"
write_pbcore_records(pbcore_writer_class, records, fasta_file_name)
log.debug("Writing ContigSet XML to {f}".format(f=file_name))
ds = ContigSet(fasta_file_name)
ds.write(file_name)
def makeReport(inReadsFN, inSummaryFN, outDir):
"""
Generate a report with ID, tables, attributes and plot groups.
inReadsFN --- an input FASTA file which has all consensus
isoforms produced by pbtranscript.py cluster.
This file is required to plot a read length histogram as part of
the report:
consensus_isoforms_readlength_hist.png
inSummaryFN --- a summary TXT file with cluster attributes,
including two attributes:
number of consensus isoforms
average length of consensus isoforms
Attributes of the report are extracted from this file.
"""
log.info("Plotting read length histogram from file: {f}".
format(f=inReadsFN))
# Collect read lengths of
reader = ContigSet(inReadsFN)
rs = [len(r.sequence) for r in reader]
reader.close()
readlengths = np.array(rs)
# Plot read length histogram
readlength_plot = create_readlength_plot(readlengths, outDir)
readlength_group = PlotGroup(Constants.PG_READLENGTH,
plots=[readlength_plot],
thumbnail=readlength_plot.thumbnail)
log.info("Plotting summary attributes from file: {f}".
format(f=inSummaryFN))
# Produce attributes based on summary.
dataset_uuids = [ContigSet(inReadsFN).uuid]
attributes = _report_to_attributes(inSummaryFN)
r = load_report_from_json(inSummaryFN)
# FIXME(nechols)(2016-03-22) not using the dataset UUIDs from these
# reports; should we be?
table = attributesToTable(attributes)
log.info(str(table))
# A report is consist of ID, tables, attributes, and plotgroups.
report = Report(Constants.R_ID,
title=meta_rpt.title,
attributes=attributes,
plotgroups=[readlength_group],
dataset_uuids=dataset_uuids)
return meta_rpt.apply_view(report)
def run_after(self, rtc, output_dir):
json_file = rtc.task.output_files[0]
chunks = load_pipeline_chunks_from_json(json_file)
n_rec = 0
with ContigSet(self.INPUT_FILES[0]) as f:
n_rec = len(f)
n_rec_chunked = 0
for chunk in chunks:
d = chunk.chunk_d
with ContigSet(d['$chunk.contigset_id']) as cs:
n_rec_chunked += len([r for r in cs])
self._check_unchunked_files(d)
self.assertEqual(n_rec_chunked, n_rec)
def test_merged_contigset(self):
fn = tempfile.NamedTemporaryFile(suffix=".contigset.xml").name
with ContigSet(upstreamData.getLambdaFasta(),
upstreamData.getFasta()) as cset:
assert len(list(cset)) == 49
assert len(cset) == 49
cset.consolidate()
cset.write(fn)
log.debug("Writing to {f}".format(f=fn))
assert len(list(cset)) == 49
assert len(cset) == 49
with ContigSet(fn) as cset:
assert len(list(cset)) == 49
assert len(cset) == 49
def test_missing_fai_error_message(self):
outdir = tempfile.mkdtemp(suffix="dataset-unittest")
inFas = os.path.join(outdir, 'infile.fasta')
# copy fasta reference to hide fai and ensure FastaReader is used
shutil.copyfile(
ReferenceSet(data.getXml(8)).toExternalFiles()[0], inFas)
rs1 = ContigSet(inFas)
with pytest.raises(IOError) as cm:
rs1.assertIndexed()
assert str(cm) == (
"Companion FASTA index (.fai) file not found or malformatted! "
"Use 'samtools faidx' to generate FASTA index.")
def test_contigset_consolidate_int_names(self):
#build set to merge
outdir = tempfile.mkdtemp(suffix="dataset-unittest")
inFas = os.path.join(outdir, 'infile.fasta')
outFas1 = os.path.join(outdir, 'tempfile1.fasta')
outFas2 = os.path.join(outdir, 'tempfile2.fasta')
# copy fasta reference to hide fai and ensure FastaReader is used
backticks('cp {i} {o}'.format(
i=ReferenceSet(data.getXml(9)).toExternalFiles()[0],
o=inFas))
rs1 = ContigSet(inFas)
double = 'B.cereus.1'
exp_double = rs1.get_contig(double)
# todo: modify the names first:
with FastaWriter(outFas1) as writer:
writer.writeRecord('5141', exp_double.sequence)
with FastaWriter(outFas2) as writer:
writer.writeRecord('5142', exp_double.sequence)
exp_double_seqs = [exp_double.sequence, exp_double.sequence]
exp_names = ['5141', '5142']
obs_file = ContigSet(outFas1, outFas2)
log.debug(obs_file.toExternalFiles())
obs_file.consolidate()
log.debug(obs_file.toExternalFiles())
# open obs and compare to exp
for name, seq in zip(exp_names, exp_double_seqs):
self.assertEqual(obs_file.get_contig(name).sequence[:], seq)
def test_contigset_write(self):
fasta = upstreamData.getLambdaFasta()
ds = ContigSet(fasta)
assert isinstance(ds.resourceReaders()[0], IndexedFastaReader)
outdir = tempfile.mkdtemp(suffix="dataset-unittest")
outfn = os.path.join(outdir, 'test.fasta')
w = FastaWriter(outfn)
for rec in ds:
w.writeRecord(rec)
w.close()
fas = FastaReader(outfn)
for rec in fas:
# make sure a __repr__ didn't slip through:
assert not rec.sequence.startswith('<')
def test_len_fastq(self):
fn = ('/pbi/dept/secondary/siv/testdata/SA3-RS/'
'lambda/2590980/0008/Analysis_Results/'
'm141115_075238_ethan_c100699872550000001'
'823139203261572_s1_p0.1.subreads.fastq')
fq_out = tempfile.NamedTemporaryFile(suffix=".fastq").name
with open(fq_out, 'w') as fqh:
with open(fn, 'r') as fih:
for line in itertools.islice(fih, 24):
fqh.write(line)
cset = ContigSet(fq_out)
assert not cset.isIndexed
assert isinstance(cset.resourceReaders()[0], FastqReader)
assert sum(1 for _ in cset) == sum(1 for _ in FastqReader(fq_out))
assert sum(1 for _ in cset) == 6
def test_contigset_write(self):
fasta = upstreamData.getLambdaFasta()
ds = ContigSet(fasta)
self.assertTrue(isinstance(ds.resourceReaders()[0],
IndexedFastaReader))
outdir = tempfile.mkdtemp(suffix="dataset-unittest")
outfn = os.path.join(outdir, 'test.fasta')
w = FastaWriter(outfn)
for rec in ds:
w.writeRecord(rec)
w.close()
fas = FastaReader(outfn)
for rec in fas:
# make sure a __repr__ didn't slip through:
self.assertFalse(rec.sequence.startswith('<'))
def test_missing_fai_error_message(self):
outdir = tempfile.mkdtemp(suffix="dataset-unittest")
inFas = os.path.join(outdir, 'infile.fasta')
# copy fasta reference to hide fai and ensure FastaReader is used
backticks('cp {i} {o}'.format(i=ReferenceSet(
data.getXml(9)).toExternalFiles()[0],
o=inFas))
rs1 = ContigSet(inFas)
with self.assertRaises(IOError) as cm:
rs1.assertIndexed()
self.assertEqual(
str(cm.exception),
("Companion FASTA index (.fai) file not found or malformatted! "
"Use 'samtools faidx' to generate FASTA index."))
def run_fasta_to_reference(input_file_name, output_file_name,
organism, reference_name,
ploidy):
"""Copied from pbcoretools/tasks/converters.py:run_fasta_to_reference()
"""
ds_in = ContigSet(input_file_name)
if len(ds_in.externalResources) > 1:
raise TypeError("Only a single FASTA file is supported as input.")
fasta_file_name = ds_in.externalResources[0].resourceId
output_dir_name = op.dirname(output_file_name)
args = [
"fasta-to-reference",
"--organism", organism,
"--ploidy", ploidy,
"--debug",
fasta_file_name,
output_dir_name,
reference_name
]
log.info(" ".join(args))
system(" ".join(args))
ref_file = op.join(output_dir_name, reference_name, "referenceset.xml")
assert op.isfile(ref_file)
with ReferenceSet(ref_file, strict=True) as ds_ref:
ds_ref.makePathsAbsolute()
log.info("saving final ReferenceSet to {f!r}".format(f=output_file_name))
ds_ref.write(output_file_name)
def run_fasta_to_reference(input_file_name,
output_file_name,
organism=None,
reference_name=None,
ploidy="haploid"):
if reference_name is None or reference_name == "":
reference_name = op.splitext(op.basename(input_file_name))[0]
ds_in = ContigSet(input_file_name)
if len(ds_in.externalResources) > 1:
raise TypeError("Only a single FASTA file is supported as input.")
fasta_file_name = ds_in.externalResources[0].resourceId
output_dir_name = op.dirname(output_file_name)
args = [
"fasta-to-reference", "--organism",
str(organism) if organism != "" else "unknown", "--ploidy",
str(ploidy) if ploidy != "" else "unknown", "--debug", fasta_file_name,
output_dir_name, reference_name
]
log.info(" ".join(args))
result = run_cmd(" ".join(args),
stdout_fh=sys.stdout,
stderr_fh=sys.stderr)
if result.exit_code != 0:
return result.exit_code
ref_file = op.join(output_dir_name, reference_name, "referenceset.xml")
assert op.isfile(ref_file)
with ReferenceSet(ref_file, strict=True) as ds_ref:
ds_ref.makePathsAbsolute()
log.info("saving final ReferenceSet to {f}".format(f=output_file_name))
ds_ref.write(output_file_name)
return 0
def getMetrics(cls):
super(TestIsoSeqCluster, cls).getMetrics()
cls.hq_fasta_file = cls.lq_fasta_file = None
for file_id, file_info in cls.datastore.get_file_dict().iteritems():
if file_info.file_type_id == FileTypes.DS_CONTIG.file_type_id:
file_name = op.basename(file_info.path)
if file_name.startswith("hq_isoforms"):
cls.hq_fasta_file = file_info.path
with ContigSet(cls.hq_fasta_file) as ds:
n = len(ds)
cls.metric_dict["num_polished_hq_isoforms_fasta"] = n
elif file_name.startswith("lq_isoforms"):
cls.lq_fasta_file = file_info.path
with ContigSet(cls.lq_fasta_file) as ds:
n = len(ds)
cls.metric_dict["num_polished_lq_isoforms_fasta"] = n
def test_len_fastq(self):
fn = ('/pbi/dept/secondary/siv/testdata/SA3-RS/'
'lambda/2590980/0008/Analysis_Results/'
'm141115_075238_ethan_c100699872550000001'
'823139203261572_s1_p0.1.subreads.fastq')
fq_out = tempfile.NamedTemporaryFile(suffix=".fastq").name
with open(fq_out, 'w') as fqh:
with open(fn, 'r') as fih:
for line in itertools.islice(fih, 24):
fqh.write(line)
cset = ContigSet(fq_out)
self.assertFalse(cset.isIndexed)
self.assertTrue(isinstance(cset.resourceReaders()[0], FastqReader))
self.assertEqual(sum(1 for _ in cset),
sum(1 for _ in FastqReader(fq_out)))
self.assertEqual(sum(1 for _ in cset), 6)
def test_missing_fai_error_message(self):
outdir = tempfile.mkdtemp(suffix="dataset-unittest")
inFas = os.path.join(outdir, 'infile.fasta')
# copy fasta reference to hide fai and ensure FastaReader is used
backticks('cp {i} {o}'.format(
i=ReferenceSet(data.getXml(9)).toExternalFiles()[0],
o=inFas))
rs1 = ContigSet(inFas)
with self.assertRaises(IOError) as cm:
rs1.assertIndexed()
self.assertEqual(
str(cm.exception),
( "Companion FASTA index (.fai) file not found or malformatted! "
"Use 'samtools faidx' to generate FASTA index."))
def test_write_contigset_records(self):
records = [FastaRecord("chr1", "acgt"), FastaRecord("chr2", "tgca")]
tmp_contigs = tempfile.NamedTemporaryFile(suffix=".contigset.xml").name
write_contigset_records(FastaWriter, records, tmp_contigs)
with ContigSet(tmp_contigs) as ds_in:
rec2 = [(rec.id, rec.sequence) for rec in ds_in]
assert rec2 == [("chr1", "acgt"), ("chr2", "tgca")]
def contigset_to_fasta(rtc):
with ContigSet(rtc.task.input_files[0]) as ds_in:
if len(ds_in.externalResources) != 1:
raise ValueError("This task assumes that the ContigSet contains "+
"only a single FASTA file.")
file_name = ds_in.externalResources[0].resourceId
os.symlink(file_name, rtc.task.output_files[0])
return 0
def as_contigset(fasta_file, xml_file):
if fasta_file == xml_file or xml_file is None:
if not op.isfile(fasta_file) or op.getsize(fasta_file) == 0:
return ContigSet()
return ContigSet(fasta_file)
file_size = op.getsize(fasta_file)
fai_file = fasta_file + ".fai"
if op.exists(fai_file):
os.remove(fai_file)
ds = ContigSet(fasta_file, generateIndices=True)
ds.write(xml_file)
if not file_size > 0:
with open(fai_file, "w") as fai:
fai.write("")
return ds
def test_contigset_consolidate_genomic_consensus(self):
"""
Verify that the contigs output by GenomicConsensus (e.g. quiver) can
be consolidated.
"""
FASTA1 = (
"lambda_NEB3011_0_60",
"GGGCGGCGACCTCGCGGGTTTTCGCTATTTATGAAAATTTTCCGGTTTAAGGCGTTTCCG")
FASTA2 = (
"lambda_NEB3011_120_180",
"CACTGAATCATGGCTTTATGACGTAACATCCGTTTGGGATGCGACTGCCACGGCCCCGTG")
FASTA3 = (
"lambda_NEB3011_60_120",
"GTGGACTCGGAGCAGTTCGGCAGCCAGCAGGTGAGCCGTAATTATCATCTGCGCGGGCGT")
files = []
for i, (header, seq) in enumerate([FASTA1, FASTA2, FASTA3]):
_files = []
for suffix in ["", "|quiver", "|plurality", "|arrow", "|poa"]:
tmpfile = tempfile.NamedTemporaryFile(suffix=".fasta").name
with open(tmpfile, "w") as f:
f.write(">{h}{s}\n{q}".format(h=header, s=suffix, q=seq))
_files.append(tmpfile)
files.append(_files)
for i in range(3):
ds = ContigSet(*[f[i] for f in files])
out1 = tempfile.NamedTemporaryFile(suffix=".contigset.xml").name
fa1 = tempfile.NamedTemporaryFile(suffix=".fasta").name
ds.consolidate(fa1)
ds.write(out1)
with ContigSet(out1) as ds_new:
assert len([rec for rec in ds_new]) == 1
def test_contigset_empty(self):
fa_file = tempfile.NamedTemporaryFile(suffix=".fasta").name
ds_file = tempfile.NamedTemporaryFile(suffix=".contigset.xml").name
open(fa_file, "w").write("")
ds = ContigSet(fa_file, strict=False)
ds.write(ds_file)
fai_file = fa_file + ".fai"
open(fai_file, "w").write("")
ds = ContigSet(fa_file, strict=True)
ds.write(ds_file)
self.assertEqual(len(ds), 0)
def test_contigset_empty(self):
fa_file = tempfile.NamedTemporaryFile(suffix=".fasta")
ds_file = tempfile.NamedTemporaryFile(suffix=".contigset.xml")
open(fa_file.name, "w").write("")
ds = ContigSet(fa_file.name, strict=False)
ds.write(ds_file.name)
fai_file = fa_file.name + ".fai"
open(fai_file, "w").write("")
ds = ContigSet(fa_file.name, strict=True)
ds.write(ds_file.name)
assert len(ds) == 0
fa_file.close()
ds_file.close()
def run_after(self, rtc, output_dir):
rpt = None
uuids = []
for file_name in rtc.task.output_files:
if file_name.endswith(".json"):
rpt = load_report_from_json(file_name)
elif file_name.endswith(".xml"):
uuids.append(ContigSet(file_name, strict=True).uuid)
else:
assert file_name.endswith(".csv")
self.assertEqual(sorted(rpt._dataset_uuids), sorted(uuids))
def gather_contigset(input_files, output_file, new_resource_file=None,
skip_empty=True):
"""
:param input_files: List of file paths
:param output_file: File Path
:param new_resource_file: the path of the file to which the other contig
files are consolidated
:param skip_empty: Ignore empty files (doesn't do much yet)
:return: Output file
:rtype: str
"""
tbr = ContigSet(*input_files)
if not new_resource_file:
if output_file.endswith('xml'):
new_resource_file = output_file[:-3] + 'fasta'
tbr.consolidate(new_resource_file)
tbr.write(output_file)
return output_file
def test_fastq_consolidate(self):
fn = ('/pbi/dept/secondary/siv/testdata/SA3-RS/'
'lambda/2590980/0008/Analysis_Results/'
'm141115_075238_ethan_c100699872550000001'
'823139203261572_s1_p0.1.subreads.fastq')
fq_out = tempfile.NamedTemporaryFile(suffix=".fastq").name
cfq_out = tempfile.NamedTemporaryFile(suffix=".fastq").name
with open(fq_out, 'w') as fqh:
with open(fn, 'r') as fih:
for line in itertools.islice(fih, 240):
fqh.write(line)
cset = ContigSet(fq_out)
cset_l = sum(1 for _ in cset)
self.assertEqual(cset_l, 60)
cset.filters.addRequirement(length=[('>', 1000)])
cset_l = sum(1 for _ in cset)
self.assertEqual(cset_l, 23)
cset.consolidate(cfq_out)
cset_l = sum(1 for _ in cset)
cfq = FastqReader(cfq_out)
self.assertEqual(cset_l, 23)
self.assertEqual(cset_l, sum(1 for _ in cfq))
def test_not_all_lowercase_contigs(self):
"""
Test that no contigs in the output ContigSet have only lowercase
characters.
"""
n_uppercase = 0
for file_id, file_info in self.datastore.get_file_dict().iteritems():
if file_info.file_type_id == FileTypes.DS_CONTIG.file_type_id:
if not file_info.is_chunked:
with ContigSet(file_info.path) as contigs:
for contig in contigs:
c = Counter(contig.sequence)
n_uppercase += c['A'] + c['C'] + c['G'] + c['T']
self.assertTrue(n_uppercase > 0, "All contigs are lowercase-only")
def _open_files(self, *input_filenames):
"""Open file handers and return."""
readers = []
for fn in input_filenames:
if ContigSetReaderWrapper.get_file_type(fn) == "FASTA":
readers.append(FastaReader(fn))
elif ContigSetReaderWrapper.get_file_type(fn) == "FASTQ":
readers.append(FastqReader(fn))
elif ContigSetReaderWrapper.get_file_type(fn) == "CONTIGSET":
readers.append(ContigSet(fn))
else:
raise IOError(
"Could not read %s as FASTA/FASTQ/CONTIGSET file." % fn)
return readers
def write_cluster_summary(summary_fn, isoforms_fa, hq_fa=None, lq_fa=None):
"""Extract number of consensus isoforms predicted, and total
number of bases in all consensuus isoforms from isoforms_fa and write
the two attributes to summary_fn.
if hq_fa (polished high-quality isoforms) is not None, report
the number of polished hq clusters
if lq_fa (polished high-quality isoforms) is not None, report
the number of polished hq clusters
"""
try:
summary = ClusterSummary()
dataset_uuids = []
with ContigSet(isoforms_fa) as reader:
for r in reader:
summary.num_consensus_isoforms += 1
summary.num_total_bases += len(r.sequence[:])
dataset_uuids.append(reader.uuid)
if hq_fa is not None and op.getsize(hq_fa) > 0:
summary.num_polished_hq_isoforms = 0
with ContigSet(hq_fa) as reader:
for r in reader:
summary.num_polished_hq_isoforms += 1
dataset_uuids.append(reader.uuid)
if lq_fa is not None and op.getsize(lq_fa) > 0:
summary.num_polished_lq_isoforms = 0
with ContigSet(lq_fa) as reader:
for r in reader:
summary.num_polished_lq_isoforms += 1
dataset_uuids.append(reader.uuid)
summary.write(summary_fn, dataset_uuids=dataset_uuids)
except ZeroDivisionError:
errMsg = "No consensus isoforms predicted."
logging.error(errMsg)
raise RuntimeError(errMsg)
def test_contigset_empty(self):
fa_file = tempfile.NamedTemporaryFile(suffix=".fasta").name
ds_file = tempfile.NamedTemporaryFile(suffix=".contigset.xml").name
open(fa_file, "w").write("")
ds = ContigSet(fa_file, strict=False)
ds.write(ds_file)
fai_file = fa_file + ".fai"
open(fai_file, "w").write("")
ds = ContigSet(fa_file, strict=True)
ds.write(ds_file)
self.assertEqual(len(ds), 0)
def n_reads_in_contigset(contigset_file):
"""Return number of reads in a contigset"""
cs = ContigSet(contigset_file)
cs.assertIndexed()
return int(cs.numRecords)
def test_empty_fastq_consolidate(self):
fn = ('/pbi/dept/secondary/siv/testdata/SA3-RS/'
'lambda/2590980/0008/Analysis_Results/'
'm141115_075238_ethan_c100699872550000001'
'823139203261572_s1_p0.1.subreads.fastq')
fq1_out = tempfile.NamedTemporaryFile(suffix="1.fastq").name
fq2_out = tempfile.NamedTemporaryFile(suffix="2.fastq").name
cfq_out = tempfile.NamedTemporaryFile(suffix=".fastq").name
# Two full
with open(fq1_out, 'w') as fqh:
with open(fn, 'r') as fih:
for line in itertools.islice(fih, 240):
fqh.write(line)
with open(fq2_out, 'w') as fqh:
with open(fn, 'r') as fih:
for line in itertools.islice(fih, 240, 480):
fqh.write(line)
cset = ContigSet(fq1_out, fq2_out)
cset_l = sum(1 for _ in cset)
self.assertEqual(cset_l, 120)
cset.consolidate(cfq_out)
cset_l = sum(1 for _ in cset)
cfq = FastqReader(cfq_out)
self.assertEqual(cset_l, 120)
self.assertEqual(cset_l, sum(1 for _ in cfq))
# one full one empty
with open(fq1_out, 'w') as fqh:
with open(fn, 'r') as fih:
for line in itertools.islice(fih, 240):
fqh.write(line)
with open(fq2_out, 'w') as fqh:
with open(fn, 'r') as fih:
fqh.write("")
cset = ContigSet(fq1_out, fq2_out)
cset_l = sum(1 for _ in cset)
self.assertEqual(cset_l, 60)
cset.consolidate(cfq_out)
cset_l = sum(1 for _ in cset)
cfq = FastqReader(cfq_out)
self.assertEqual(cset_l, 60)
self.assertEqual(cset_l, sum(1 for _ in cfq))
# one empty one full
with open(fq1_out, 'w') as fqh:
with open(fn, 'r') as fih:
fqh.write("")
with open(fq2_out, 'w') as fqh:
with open(fn, 'r') as fih:
for line in itertools.islice(fih, 240):
fqh.write(line)
cset = ContigSet(fq1_out, fq2_out)
cset_l = sum(1 for _ in cset)
self.assertEqual(cset_l, 60)
cset.consolidate(cfq_out)
cset_l = sum(1 for _ in cset)
cfq = FastqReader(cfq_out)
self.assertEqual(cset_l, 60)
self.assertEqual(cset_l, sum(1 for _ in cfq))
# both empty
with open(fq1_out, 'w') as fqh:
with open(fn, 'r') as fih:
fqh.write("")
with open(fq2_out, 'w') as fqh:
with open(fn, 'r') as fih:
fqh.write("")
cset = ContigSet(fq1_out, fq2_out)
cset_l = sum(1 for _ in cset)
self.assertEqual(cset_l, 0)
cset.consolidate(cfq_out)
cset_l = sum(1 for _ in cset)
cfq = FastqReader(cfq_out)
self.assertEqual(cset_l, 0)
self.assertEqual(cset_l, sum(1 for _ in cfq))
def test_contigset_consolidate(self):
#build set to merge
outdir = tempfile.mkdtemp(suffix="dataset-unittest")
inFas = os.path.join(outdir, 'infile.fasta')
outFas1 = os.path.join(outdir, 'tempfile1.fasta')
outFas2 = os.path.join(outdir, 'tempfile2.fasta')
# copy fasta reference to hide fai and ensure FastaReader is used
backticks('cp {i} {o}'.format(
i=ReferenceSet(data.getXml(9)).toExternalFiles()[0],
o=inFas))
rs1 = ContigSet(inFas)
singletons = ['A.baumannii.1', 'A.odontolyticus.1']
double = 'B.cereus.1'
reader = rs1.resourceReaders()[0]
exp_double = rs1.get_contig(double)
exp_singles = [rs1.get_contig(name) for name in singletons]
# todo: modify the names first:
with FastaWriter(outFas1) as writer:
writer.writeRecord(exp_singles[0])
writer.writeRecord(exp_double.name + '_10_20', exp_double.sequence)
with FastaWriter(outFas2) as writer:
writer.writeRecord(exp_double.name + '_0_10',
exp_double.sequence + 'ATCGATCGATCG')
writer.writeRecord(exp_singles[1])
exp_double_seq = ''.join([exp_double.sequence,
'ATCGATCGATCG',
exp_double.sequence])
exp_single_seqs = [rec.sequence for rec in exp_singles]
acc_file = ContigSet(outFas1, outFas2)
acc_file.induceIndices()
log.debug(acc_file.toExternalFiles())
self.assertEqual(len(acc_file), 4)
self.assertEqual(len(list(acc_file)), 4)
acc_file.consolidate()
log.debug(acc_file.toExternalFiles())
# open acc and compare to exp
for name, seq in zip(singletons, exp_single_seqs):
self.assertEqual(acc_file.get_contig(name).sequence[:], seq)
self.assertEqual(acc_file.get_contig(double).sequence[:],
exp_double_seq)
self.assertEqual(len(acc_file._openReaders), 1)
self.assertEqual(len(acc_file.index), 3)
self.assertEqual(len(acc_file._indexMap), 3)
self.assertEqual(len(acc_file), 3)
self.assertEqual(len(list(acc_file)), 3)
def consolidateXml(args):
"""Combine BAMs and apply the filters described in the XML file, producing
one consolidated XML"""
dset = ContigSet(args.infile)
dset.consolidate(args.datafile)
dset.write(args.xmlfile)
def makeReport(inReadsFN, hq_isoforms_fq, lq_isoforms_fq, inSummaryFN, outDir):
"""
Generate a report with ID, tables, attributes and plot groups.
inReadsFN --- an input FASTA file which has all consensus
isoforms produced by pbtranscript.py cluster.
This file is required to plot a read length histogram as part of
the report:
consensus_isoforms_readlength_hist.png
hq_isoforms_fq/lq_isoforms_lq --- input FASTQ files which has
all HQ/LQ isoforms produced by pbtranscript.py cluster.
These two files will be required to plot the average QV histograms:
hq_lq_isoforms_avgqv_hist.png
inSummaryFN --- a summary TXT file with cluster attributes,
including two attributes:
number of consensus isoforms
average length of consensus isoforms
Attributes of the report are extracted from this file.
"""
log.info("Plotting read length histogram from file: {f}".
format(f=inReadsFN))
# Collect read lengths of
reader = ContigSet(inReadsFN)
rs = [len(r.sequence) for r in reader]
reader.close()
readlengths = np.array(rs).astype(float)
# Plot read length histogram
readlength_plot = create_readlength_plot(readlengths, outDir)
readlength_group = PlotGroup(Constants.PG_READLENGTH,
plots=[readlength_plot],
thumbnail=readlength_plot.thumbnail)
# Collect average qvs
hq_qvs = [np.mean(r.quality) for r in ContigSet(hq_isoforms_fq)]
lq_qvs = [np.mean(r.quality) for r in ContigSet(lq_isoforms_fq)]
avgqvs = np.array(hq_qvs + lq_qvs)
# Plot average qv histogram
avgqv_plot = create_avgqv_plot(avgqvs, outDir)
avgqv_group = PlotGroup(Constants.PG_AVGQV,
plots=[avgqv_plot],
thumbnail=avgqv_plot.thumbnail)
log.info("Plotting summary attributes from file: {f}".
format(f=inSummaryFN))
# Produce attributes based on summary.
dataset_uuids = [ContigSet(inReadsFN).uuid]
attributes = _report_to_attributes(inSummaryFN)
r = load_report_from_json(inSummaryFN)
# FIXME(nechols)(2016-03-22) not using the dataset UUIDs from these
# reports; should we be?
table = attributesToTable(attributes)
log.info(str(table))
# A report is consist of ID, tables, attributes, and plotgroups.
report = Report(Constants.R_ID,
attributes=attributes,
plotgroups=[readlength_group, avgqv_group],
dataset_uuids=dataset_uuids)
return spec.apply_view(report)
