def deplete_blastn(inFastq, outFastq, refDbs) :
'Use blastn to remove reads that match at least one of the databases.'
## Get tools
noBlastHits_v3Path = os.path.join(util.file.get_scripts_path(),
'noBlastHits_v3.py')
## Convert to fasta
inFasta = mkstempfname('.fasta')
read_utils.fastq_to_fasta(inFastq, inFasta)
## Run blastn using each of the databases in turn
blastOutFiles = []
for db in refDbs :
log.info("running blastn on %s against %s", inFastq, db)
blastOutFiles += blastn_chunked_fasta(inFasta, db)
## Combine results from different databases
blastOutCombined = mkstempfname('.txt')
catCmd = ['cat'] + blastOutFiles
log.debug(' '.join(catCmd) + '> ' + blastOutCombined)
with open(blastOutCombined, 'wt') as outf:
subprocess.check_call(catCmd, stdout = outf)
## run noBlastHits_v3.py to extract reads with no blast hits
# TODO: slurp the small amount of code in this script into here
noBlastHitsCmd = ['python', noBlastHits_v3Path, '-b', blastOutCombined,
'-r', inFastq, '-m', 'nohit']
log.debug(' '.join(noBlastHitsCmd) + '> ' + outFastq)
with util.file.open_or_gzopen(outFastq, 'wt') as outf :
subprocess.check_call(noBlastHitsCmd, stdout = outf)
def deplete_blastn_bam(inBam, db, outBam, threads, chunkSize=1000000, JVMmemory=None):
'Use blastn to remove reads that match at least one of the databases.'
#blastnPath = tools.blast.BlastnTool().install_and_get_path()
fastq1 = mkstempfname('.1.fastq')
fastq2 = mkstempfname('.2.fastq')
fasta = mkstempfname('.1.fasta')
blast_hits = mkstempfname('.blast_hits.txt')
halfBam = mkstempfname('.half.bam')
blastOutFile = mkstempfname('.hits.txt')
# Initial BAM -> FASTQ pair
tools.samtools.SamtoolsTool().bam2fq(inBam, fastq1, fastq2)
# Find BLAST hits against FASTQ1
read_utils.fastq_to_fasta(fastq1, fasta)
os.unlink(fastq1)
os.unlink(fastq2)
log.info("running blastn on %s pair 1 against %s", inBam, db)
blastOutFiles = blastn_chunked_fasta(fasta, db, chunkSize, threads)
with open(blast_hits, 'wt') as outf:
for blastOutFile in blastOutFiles:
with open(blastOutFile, 'rt') as inf:
for line in inf:
idVal = line.split('\t')[0].strip()
if idVal.endswith('/1') or idVal.endswith('/2'):
idVal = idVal[:-2]
outf.write(idVal + '\n')
os.unlink(blastOutFile)
# Deplete BAM of hits in FASTQ1
tools.picard.FilterSamReadsTool().execute(inBam, True, blast_hits, halfBam, JVMmemory=JVMmemory)
# Depleted BAM -> FASTQ pair
tools.samtools.SamtoolsTool().bam2fq(halfBam, fastq1, fastq2)
# Find BLAST hits against FASTQ2 (which is already smaller than before)
read_utils.fastq_to_fasta(fastq2, fasta)
os.unlink(fastq1)
os.unlink(fastq2)
log.info("running blastn on %s pair 2 against %s", inBam, db)
blastOutFiles = blastn_chunked_fasta(fasta, db, chunkSize, threads)
with open(blast_hits, 'wt') as outf:
for blastOutFile in blastOutFiles:
with open(blastOutFile, 'rt') as inf:
for line in inf:
idVal = line.split('\t')[0].strip()
if idVal.endswith('/1') or idVal.endswith('/2'):
idVal = idVal[:-2]
outf.write(idVal + '\n')
os.unlink(blastOutFile)
# Deplete BAM of hits against FASTQ2
tools.picard.FilterSamReadsTool().execute(halfBam, True, blast_hits, outBam, JVMmemory=JVMmemory)
# Clean up
for fn in (fasta, blast_hits, halfBam):
os.unlink(fn)
def deplete_blastn_bam(inBam, db, outBam, chunkSize=1000000, JVMmemory=None):
'Use blastn to remove reads that match at least one of the databases.'
#blastnPath = tools.blast.BlastnTool().install_and_get_path()
fastq1 = mkstempfname('.1.fastq')
fastq2 = mkstempfname('.2.fastq')
fasta = mkstempfname('.1.fasta')
blast_hits = mkstempfname('.blast_hits.txt')
halfBam = mkstempfname('.half.bam')
blastOutFile = mkstempfname('.hits.txt')
# Initial BAM -> FASTQ pair
tools.picard.SamToFastqTool().execute(inBam, fastq1, fastq2)
# Find BLAST hits against FASTQ1
read_utils.fastq_to_fasta(fastq1, fasta)
os.unlink(fastq1)
os.unlink(fastq2)
log.info("running blastn on %s pair 1 against %s", inBam, db)
blastOutFiles = blastn_chunked_fasta(fasta, db, chunkSize)
with open(blast_hits, 'wt') as outf:
for blastOutFile in blastOutFiles:
with open(blastOutFile, 'rt') as inf:
for line in inf:
idVal = line.split('\t')[0].strip()
if idVal.endswith('/1') or idVal.endswith('/2'):
idVal = idVal[:-2]
outf.write(idVal + '\n')
os.unlink(blastOutFile)
# Deplete BAM of hits in FASTQ1
tools.picard.FilterSamReadsTool().execute(inBam, True, blast_hits, halfBam, JVMmemory=JVMmemory)
# Depleted BAM -> FASTQ pair
tools.picard.SamToFastqTool().execute(halfBam, fastq1, fastq2)
# Find BLAST hits against FASTQ2 (which is already smaller than before)
read_utils.fastq_to_fasta(fastq2, fasta)
os.unlink(fastq1)
os.unlink(fastq2)
log.info("running blastn on %s pair 2 against %s", inBam, db)
blastOutFiles = blastn_chunked_fasta(fasta, db, chunkSize)
with open(blast_hits, 'wt') as outf:
for blastOutFile in blastOutFiles:
with open(blastOutFile, 'rt') as inf:
for line in inf:
idVal = line.split('\t')[0].strip()
if idVal.endswith('/1') or idVal.endswith('/2'):
idVal = idVal[:-2]
outf.write(idVal + '\n')
os.unlink(blastOutFile)
# Deplete BAM of hits against FASTQ2
tools.picard.FilterSamReadsTool().execute(halfBam, True, blast_hits, outBam, JVMmemory=JVMmemory)
# Clean up
for fn in (fasta, blast_hits, halfBam):
os.unlink(fn)
def deplete_blastn(inFastq, outFastq, refDbs, threads=1, chunkSize=1000000):
'Use blastn to remove reads that match at least one of the databases.'
# Convert to fasta
inFasta = mkstempfname('.fasta')
read_utils.fastq_to_fasta(inFastq, inFasta)
# Run blastn using each of the databases in turn
blastOutFiles = []
for db in refDbs:
log.info("running blastn on %s against %s", inFastq, db)
blastOutFiles += blastn_chunked_fasta(inFasta, db, chunkSize, threads)
# Combine results from different databases
blastOutCombined = mkstempfname('.txt')
catCmd = ['cat'] + blastOutFiles
log.debug(' '.join(catCmd) + '> ' + blastOutCombined)
with open(blastOutCombined, 'wt') as outf:
subprocess.check_call(catCmd, stdout=outf)
# extract reads with no blast hits
no_blast_hits(blastOutCombined, inFastq, outFastq)
def deplete_blastn_bam(inBam,
db,
outBam,
threads,
chunkSize=1000000,
JVMmemory=None):
'Use blastn to remove reads that match at least one of the databases.'
fastq1 = mkstempfname('.1.fastq')
fasta = mkstempfname('.1.fasta')
blast_hits = mkstempfname('.blast_hits.txt')
blastOutFile = mkstempfname('.hits.txt')
# Initial BAM -> FASTQ pair
tools.samtools.SamtoolsTool().bam2fq(inBam, fastq1)
# Find BLAST hits
read_utils.fastq_to_fasta(fastq1, fasta)
os.unlink(fastq1)
log.info("running blastn on %s against %s", inBam, db)
blastOutFiles = blastn_chunked_fasta(fasta, db, chunkSize, threads)
with open(blast_hits, 'wt') as outf:
for blastOutFile in blastOutFiles:
with open(blastOutFile, 'rt') as inf:
for line in inf:
idVal = line.split('\t')[0].strip()
if idVal.endswith('/1') or idVal.endswith('/2'):
idVal = idVal[:-2]
outf.write(idVal + '\n')
os.unlink(blastOutFile)
os.unlink(fasta)
# Deplete BAM of hits
tools.picard.FilterSamReadsTool().execute(inBam,
True,
blast_hits,
outBam,
JVMmemory=JVMmemory)
os.unlink(blast_hits)
def deplete_blastn(inFastq, outFastq, refDbs, threads=1, chunkSize=1000000):
'Use blastn to remove reads that match at least one of the databases.'
# Convert to fasta
inFasta = mkstempfname('.fasta')
read_utils.fastq_to_fasta(inFastq, inFasta)
# Run blastn using each of the databases in turn
blastOutFiles = []
for db in refDbs:
log.info("running blastn on %s against %s", inFastq, db)
blastOutFiles += blastn_chunked_fasta(inFasta, db, chunkSize, threads)
# Combine results from different databases
blastOutCombined = mkstempfname('.txt')
catCmd = ['cat'] + blastOutFiles
log.debug(' '.join(catCmd) + '> ' + blastOutCombined)
with open(blastOutCombined, 'wt') as outf:
subprocess.check_call(catCmd, stdout=outf)
# extract reads with no blast hits
no_blast_hits(blastOutCombined, inFastq, outFastq)
def deplete_blastn(inFastq, outFastq, refDbs) :
'Use blastn to remove reads that match at least one of the databases.'
## Get tools
blastnPath = tools.blast.BlastnTool().install_and_get_path()
noBlastHits_v3Path = os.path.join(util.file.get_scripts_path(),
'noBlastHits_v3.py')
## Convert to fasta
inFasta = mkstempfname('.fasta')
read_utils.fastq_to_fasta(inFastq, inFasta)
## Run blastn using each of the databases in turn
blastOutFiles = [mkstempfname() for db in refDbs]
for db, blastOutFile in zip(refDbs, blastOutFiles) :
log.info("running blastn on {} against {}".format(inFastq, db))
blastnCmd = [blastnPath, '-db', db,
'-word_size', '16', '-evalue', '1e-6', '-outfmt', '6',
'-num_descriptions', '2', '-num_alignments', '2',
'-query', inFasta, '-out', blastOutFile]
log.debug(' '.join(blastnCmd))
subprocess.check_call(blastnCmd)
## Combine results from different databases
blastOutCombined = mkstempfname('.txt')
catCmd = ['cat'] + blastOutFiles
log.debug(' '.join(catCmd) + '> ' + blastOutCombined)
with open(blastOutCombined, 'wt') as outf:
subprocess.check_call(catCmd, stdout = outf)
## run noBlastHits_v3.py to extract reads with no blast hits
# TODO: slurp the small amount of code in this script into here
noBlastHitsCmd = ['python', noBlastHits_v3Path, '-b', blastOutCombined,
'-r', inFastq, '-m', 'nohit']
log.debug(' '.join(noBlastHitsCmd) + '> ' + outFastq)
with util.file.open_or_gzopen(outFastq, 'wt') as outf :
subprocess.check_call(noBlastHitsCmd, stdout = outf)
def deplete_blastn_bam(inBam, db, outBam):
'Use blastn to remove reads that match at least one of the databases.'
blastnPath = tools.blast.BlastnTool().install_and_get_path()
fastq1 = mkstempfname('.1.fastq')
fastq2 = mkstempfname('.2.fastq')
fasta = mkstempfname('.1.fasta')
blast_hits = mkstempfname('.blast_hits.txt')
halfBam = mkstempfname('.half.bam')
blastOutFile = mkstempfname('.hits.txt')
# Initial BAM -> FASTQ pair
tools.picard.SamToFastqTool().execute(inBam, fastq1, fastq2)
# Find BLAST hits against FASTQ1
read_utils.fastq_to_fasta(fastq1, fasta)
os.unlink(fastq1)
os.unlink(fastq2)
log.info("running blastn on {} pair 1 against {}".format(inBam, db))
blastnCmd = [blastnPath, '-db', db,
'-word_size', '16', '-evalue', '1e-6', '-outfmt', '6',
'-num_descriptions', '2', '-num_alignments', '2',
'-query', fasta, '-out', blastOutFile]
log.debug(' '.join(blastnCmd))
subprocess.check_call(blastnCmd)
with open(blast_hits, 'wt') as outf:
with open(blastOutFile, 'rt') as inf:
for line in inf:
id = line.split('\t')[0].strip()
if id.endswith('/1') or id.endswith('/2'):
id = id[:-2]
outf.write(id+'\n')
os.unlink(blastOutFile)
# Deplete BAM of hits in FASTQ1
tools.picard.FilterSamReadsTool().execute(inBam, True, blast_hits, halfBam)
# Depleted BAM -> FASTQ pair
tools.picard.SamToFastqTool().execute(halfBam, fastq1, fastq2)
# Find BLAST hits against FASTQ2 (which is already smaller than before)
read_utils.fastq_to_fasta(fastq2, fasta)
os.unlink(fastq1)
os.unlink(fastq2)
log.info("running blastn on {} pair 2 against {}".format(inBam, db))
blastnCmd = [blastnPath, '-db', db,
'-word_size', '16', '-evalue', '1e-6', '-outfmt', '6',
'-num_descriptions', '2', '-num_alignments', '2',
'-query', fasta, '-out', blastOutFile]
log.debug(' '.join(blastnCmd))
subprocess.check_call(blastnCmd)
with open(blast_hits, 'wt') as outf:
with open(blastOutFile, 'rt') as inf:
for line in inf:
id = line.split('\t')[0].strip()
if id.endswith('/1') or id.endswith('/2'):
id = id[:-2]
outf.write(id+'\n')
os.unlink(blastOutFile)
# Deplete BAM of hits against FASTQ2
tools.picard.FilterSamReadsTool().execute(halfBam, True, blast_hits, outBam)
# Clean up
map(os.unlink, (fasta, blast_hits, halfBam))
