def processSample(sampleName, sampleNameDict):
'''pipeline for sample '''
# process communication
mng = Manager()
libraryBamFileList = mng.list()
# This libraryBamFileList is a list contains seval bams from one sample.
# init mult-processer
record = []
for sampleID in sampleNameDict:
Processer = Process(name=sampleID,
target=processSampleFromLibarary,
args=(
sampleID,
sampleNameDict[sampleID],
libraryBamFileList,
))
Processer.start()
record.append(Processer)
# wait for processer
for proc in record:
proc.join()
###################### 2.1 post mapping ####################
bamFileDir = os.path.dirname(libraryBamFileList[0])
finalBamFilePath = os.path.join(bamFileDir, "%s_final.bam" % sampleName)
# merge the bam from each line to one final bam
command = NGSTools.picard_merge(libraryBamFileList, finalBamFilePath, cfg)
NGSTools.writeCommands(command,
args.outDir + '/mapping/picard_mergebam_' +
sampleName + '.sh',
run=_run)
###################### 3. romove duplicates ##################
if RMDUP:
command = NGSTools.picard_rmdup(finalBamFilePath, True, cfg)
NGSTools.writeCommands(command,
args.outDir + '/mapping/picard_rmdup_' +
sampleName + '.sh',
run=_run)
finalBamFilePath = re.sub(r'.bam$', '.rmdup.bam', finalBamFilePath)
##################### 4. DMR calling ##################
if Methylation_extractor:
NGSTools.methylation_extractor(finalBamFilePath,
bamFileDir + '/' + sampleName, cfg)
def processSample(sampleName, sampleNameDict):
'''pipeline for sample '''
# process communication
mng = Manager()
libraryBamFileList = mng.list()
# This libraryBamFileList is a list contains seval bams from one sample.
# init mult-processer
record = []
for sampleID in sampleNameDict:
Processer = Process(name = sampleID, target = processSampleFromLibarary, args = (sampleID, sampleNameDict[sampleID], libraryBamFileList, ))
Processer.start()
record.append(Processer)
# wait for processer
for proc in record:
proc.join()
###################### 2.1 post mapping ####################
bamFileDir = os.path.dirname(libraryBamFileList[0])
finalBamFilePath = os.path.join(bamFileDir, "%s_properly.bam" % sampleName)
mergedBamFilePath = os.path.join(bamFileDir, "%s_merged.bam" % sampleName)
# merge the bam from each lane to one final bam
command = NGSTools.picard_merge(libraryBamFileList, mergedBamFilePath, cfg)
NGSTools.writeCommands(command, bamFileDir+'/picard_mergebam_'+sampleName+'.sh', run=_run)
###################### 2.2. filter bam ######################
#command = 'samtools view -Sb -h -f 2 -q 10 %s > %s ' % (mergedBamFilePath, finalBamFilePath)
#command = 'samtools view -Sb -h -q 10 %s > %s ' % (mergedBamFilePath, finalBamFilePath)
#NGSTools.writeCommands(command, bamFileDir+'/filterBam_'+sampleName+'.sh', run=_run)
###################### 3. romove duplicates ##################
if RMDUP:
command = NGSTools.picard_rmdup(mergedBamFilePath, True, cfg)
NGSTools.writeCommands(command, bamFileDir+'/picard_rmdup_'+sampleName+'.sh', run=_run)
finalBamFilePath = re.sub(r'.bam$', '.rmdup.bam', finalBamFilePath)
####################### 4. call SNP ######################
if MPILEUP:
SNP_out = os.path.join(args.outDir, "SNP", sampleName)
NGSTools._mkdir(SNP_out)
command, rawVcf = NGSTools.bcftools_call(finalBamFilePath, cfg, outdir=SNP_out, sampleName=sampleName)
NGSTools.writeCommands(command, SNP_out+'/bcftools_call_'+sampleName+'.sh', run=_run)
outVcf = rawVcf.replace("vcf$", "flt.vcf")
command = NGSTools.bcftools_filter(rawVcf, outVcf, cfg)
NGSTools.writeCommands(command, SNP_out+'/bcftools_filter_'+sampleName+'.sh', run=_run)
def processSample(sampleName, sampleNameDict): '''pipeline for sample ''' # process communication mng = Manager() libraryBamFileList = mng.list() # This libraryBamFileList is a list contains seval bams from one sample. # init mult-processer record = [] for sampleID in sampleNameDict: Processer = Process(name = sampleID, target = processSampleFromLibarary, args = (sampleID, sampleNameDict[sampleID], libraryBamFileList, )) Processer.start() record.append(Processer) # wait for processer for proc in record: proc.join() ###################### 2.1 post mapping #################### bamFileDir = os.path.dirname(libraryBamFileList[0]) finalBamFilePath = os.path.join(bamFileDir, "%s_final.bam" % sampleName) # merge the bam from each line to one final bam command = NGSTools.picard_merge(libraryBamFileList, finalBamFilePath, cfg) NGSTools.writeCommands(command, args.outDir+'/mapping/picard_mergebam_'+sampleName+'.sh', run=_run) ###################### 3. romove duplicates ################## if RMDUP: command = NGSTools.picard_rmdup(finalBamFilePath, True, cfg) NGSTools.writeCommands(command, args.outDir+'/mapping/picard_rmdup_'+sampleName+'.sh', run=_run) finalBamFilePath = re.sub(r'.bam$', '.rmdup.bam', finalBamFilePath) ##################### 4. DMR calling ################## if Methylation_extractor: NGSTools.methylation_extractor(finalBamFilePath, bamFileDir+'/'+sampleName, cfg)
def processSample(sampleName, sampleNameDict):
'''pipeline for sample '''
# process communication
mng = Manager()
libraryBamFileList = mng.list()
# This libraryBamFileList is a list contains seval bams from one sample.
# init mult-processer
record = []
for sampleID in sampleNameDict:
Processer = Process(name=sampleID,
target=processSampleFromLibarary,
args=(
sampleID,
sampleNameDict[sampleID],
libraryBamFileList,
))
Processer.start()
record.append(Processer)
# wait for processer
for proc in record:
proc.join()
###################### 2.1 post mapping ####################
bamFileDir = os.path.dirname(libraryBamFileList[0])
finalBamFilePath = os.path.join(bamFileDir, "%s_properly.bam" % sampleName)
mergedBamFilePath = os.path.join(bamFileDir, "%s_merged.bam" % sampleName)
# merge the bam from each line to one final bam
command = NGSTools.picard_merge(libraryBamFileList, mergedBamFilePath, cfg)
NGSTools.writeCommands(command,
bamFileDir + '/picard_mergebam_' + sampleName +
'.sh',
run=_run)
###################### 3. filter bam ######################
#command = 'samtools view -Sb -h -f 2 -q 10 %s > %s ' % (mergedBamFilePath, finalBamFilePath)
command = 'samtools view -Sb -h -q 10 %s > %s ' % (mergedBamFilePath,
finalBamFilePath)
NGSTools.writeCommands(command,
bamFileDir + '/filterBam_' + sampleName + '.sh',
run=_run)
###################### 4. romove duplicates ##################
if RMDUP:
command = NGSTools.picard_rmdup(finalBamFilePath, True, cfg)
NGSTools.writeCommands(command,
bamFileDir + '/picard_rmdup_' + sampleName +
'.sh',
run=_run)
finalBamFilePath = re.sub(r'.bam$', '.rmdup.bam', finalBamFilePath)
def processSample(sampleName, sampleNameDict): '''pipeline for sample ''' # process communication mng = Manager() libraryBamFileList = mng.list() # This libraryBamFileList is a list contains seval bams from one sample. # init mult-processer record = [] for sampleID in sampleNameDict: Processer = Process(name = sampleID, target = processSampleFromLibarary, args = (sampleID, sampleNameDict[sampleID], libraryBamFileList, )) Processer.start() record.append(Processer) # wait for processer for proc in record: proc.join() ###################### 2.1 post mapping #################### bamFileDir = os.path.dirname(libraryBamFileList[0]) finalBamFilePath = os.path.join(bamFileDir, "%s_properly.bam" % sampleName) mergedBamFilePath = os.path.join(bamFileDir, "%s_merged.bam" % sampleName) # merge the bam from each line to one final bam command = NGSTools.picard_merge(libraryBamFileList, mergedBamFilePath, cfg) NGSTools.writeCommands(command, bamFileDir+'/picard_mergebam_'+sampleName+'.sh', run=_run) ###################### 3. filter bam ###################### command = 'samtools view -Sb -h -f 2 -q 10 %s > %s ' % (mergedBamFilePath, finalBamFilePath) NGSTools.writeCommands(command, bamFileDir+'/filterBam_'+sampleName+'.sh', run=_run) ###################### 4. romove duplicates ################## if RMDUP: command = NGSTools.picard_rmdup(finalBamFilePath, True, cfg) NGSTools.writeCommands(command, bamFileDir+'/picard_rmdup_'+sampleName+'.sh', run=_run) finalBamFilePath = re.sub(r'.bam$', '.rmdup.bam', finalBamFilePath)
def processSample(line, condition, transcripts, countsFiles, finalBam,
expressCXB):
cols = line.strip().split('\t')
if len(cols) == 3:
# single end library
fq2 = '-'
else:
# paired end
fq2 = cols[3]
sample = {
'name': cols[0],
'condition': cols[1],
'fq1': cols[2],
'fq2': fq2,
'bam': ''
}
########################## 0. init #########################
#__init__(self, sampleName, outdir, fq1, fq2='', quanlityBase='32', cfgfile='~/.NGSTools.cfg'):
mySample = NGSTools.NGSTools(sample['name'],
args.outDir,
sample['fq1'],
sample['fq2'],
libType=args.libraryType,
cfgfile=os.path.abspath(args.config))
if QC:
#################### 1. Quality Control ####################
###### 1.1 cut adapter ######
if args.dataType == 'raw':
#mySample.cutadapter(adapter5='', adapter3='AATGATACGGCGACCACCGAGATCT', run = _run)
mySample.cutadapter(run=_run)
### Nextera Kit
#mySample.cutadapter(adapter5='CTGTCTCTTATACAC', adapter3='CTGTCTCTTATACAC',run = _run)
#mySample.rm_lowQual(run = _run)
else:
pass
##### 1.2 fastqc #####
mySample.QC_fastqc(run=_run)
if Mapping:
######################## 2. Mapping ########################
sample['bam'] = mySample.tophat2(run=_run)
if condition.has_key(sample['condition']):
#condition[sample['condition']][sample['name']] = sample['bam']
condition[sample['condition']] += "," + sample['bam']
else:
#condition[sample['condition']] = {sample['name'] : sample['bam']}
condition[sample['condition']] = sample['bam']
if GFold:
# GFold count
mySample.gfoldCount(run=_run)
if DESeq:
# DESeq2
count = mySample.HTSeq_count(run=_run)
countsFiles[count] = sample['condition'] + '|' + sample['name']
if GATK:
# remove duplicates
mySample.rmdup(run=_run)
# picard reorder
mySample.picard_reorder(run=_run)
# splitN
mySample.splitN(run=_run)
# realign
realnBam = mySample.realn(run=_run)
# recal need known SNP site
# recal
#recalBam = mySample.recal(run = _run)
finalBam[realnBam] = sample['condition']
# samtools call SNP/InDel
mySample.samtools_call(run=_run)
mySample.samtools_filter(run=_run)
######################## DEGs calling preparation ########################
if Cufflinks:
##### 3. cufflinks #####
cuffdir = os.path.join(args.outDir, 'cufflinks')
if not os.path.exists(cuffdir):
os.mkdir(cuffdir)
# cufflinks #
command = 'cufflinks --library-type %s -p 4 -g %s -o %s %s' % (
args.libraryType, cfg.gtf,
os.path.join(cuffdir, sample['condition'] + '_' + sample['name']),
sample['bam'])
NGSTools.writeCommands(command,
cuffdir + '/cufflinks_%s.sh' % sample['name'],
_run)
transcripts.append(
os.path.join(cuffdir, sample['condition'] + '_' + sample['name'],
'transcripts.gtf'))
record.append(P)
for P in record:
P.join()
######################## DEGs calling ########################
if Cufflinks:
# cuffmerge #
cuffdir = os.path.join(args.outDir, 'cufflinks')
with open(cuffdir + '/assemblies.txt', 'w') as writer:
writer.write('\n'.join(transcripts))
command = 'cuffmerge -o %s -g %s -s %s -p 10 %s' % (
cuffdir + '/merged_asm', cfg.gtf, cfg.genome,
cuffdir + '/assemblies.txt')
NGSTools.writeCommands(command, cuffdir + '/cuffmerge.sh', _run)
# cuffnorm #
cuffnorm_dir = os.path.join(cuffdir, 'cuffnorm')
try:
os.mkdir(cuffnorm_dir)
except:
pass
cond = ','.join(condition.keys())
bams = ' '.join(condition.values())
command = 'mkdir cuffnorm\ncuffnorm --library-type %s -o %s -L %s %s %s' % (
args.libraryType, cuffnorm_dir, cond, cfg.gtf, bams)
NGSTools.writeCommands(command, cuffdir + '/cuffnorm.sh', _run)
# cuffdiff #
def processSample(line, condition, transcripts, countsFiles, finalBam):
cols = line.strip().split('\t')
if len(cols) == 3:
# single end library
fq2 = '-'
else:
# paired end
fq2 = cols[3]
sample = {
'name' : cols[0],
'condition' : cols[1],
'fq1' : cols[2],
'fq2' : fq2,
'bam' : ''
}
########################## 0. init #########################
#__init__(self, sampleName, outdir, fq1, fq2='', quanlityBase='32', cfgfile='~/.NGSTools.cfg'):
mySample = NGSTools.NGSTools(sample['name'], args.outDir, sample['fq1'], sample['fq2'], cfgfile=os.path.abspath(args.config))
if QC:
#################### 1. Quality Control ####################
###### 1.1 cut adapter ######
if args.dataType == 'raw':
#mySample.cutadapter(adapter5='', adapter3='AATGATACGGCGACCACCGAGATCT', run = _run)
mySample.cutadapter(run = _run)
#mySample.rm_lowQual(run = _run)
else:
pass
##### 1.2 fastqc #####
mySample.QC_fastqc(run = _run)
if Mapping:
######################## 2. Mapping ########################
sample['bam'] = mySample.tophat2(run = _run)
if condition.has_key(sample['condition']):
#condition[sample['condition']][sample['name']] = sample['bam']
condition[sample['condition']] += ","+sample['bam']
else:
#condition[sample['condition']] = {sample['name'] : sample['bam']}
condition[sample['condition']] = sample['bam']
if GFold:
# GFold count
mySample.gfoldCount(run = _run)
if DESeq2:
# DESeq2
count = mySample.HTSeq_count(run = _run)
countsFiles[count] = sample['condition']
if GATK:
# remove duplicates
mySample.rmdup(run = _run)
# picard reorder
mySample.picard_reorder(run = _run)
# splitN
mySample.splitN(run = _run)
# realign
realnBam = mySample.realn(run = _run)
# recal need known SNP site
# recal
#recalBam = mySample.recal(run = _run)
finalBam[realnBam] = sample['condition']
# samtools call SNP/InDel
mySample.samtools_call(run = _run)
mySample.samtools_filter(run = _run)
######################## DEGs calling preparation ########################
if Cufflinks:
##### 3. cufflinks #####
cuffdir = os.path.join(args.outDir, 'cufflinks')
if not os.path.exists(cuffdir):
os.mkdir(cuffdir)
# cufflinks #
command = 'cufflinks -p 4 -g %s -o %s %s' % (cfg.gtf, os.path.join(cuffdir, sample['condition']+'_'+sample['name']), sample['bam'])
NGSTools.writeCommands(command, cuffdir+'/cufflinks_%s.sh' % sample['name'], _run)
transcripts.append(os.path.join(cuffdir, sample['condition']+'_'+sample['name'], 'transcripts.gtf'))
record.append(P)
for P in record:
P.join()
######################## DEGs calling ########################
if Cufflinks:
# cuffmerge #
cuffdir = os.path.join(args.outDir, 'cufflinks')
with open(cuffdir+'/assemblies.txt', 'w') as writer:
writer.write('\n'.join(transcripts))
command = 'cuffmerge -o %s -g %s -s %s -p 10 %s' % (cuffdir+'/merged_asm', cfg.gtf, cfg.genome, cuffdir+'/assemblies.txt')
NGSTools.writeCommands(command, cuffdir+'/cuffmerge.sh', _run)
# cuffdiff #
if len(condition) != 2:
print 'WARNING: condition'
command = 'cuffdiff -o %s -b %s -p 10 -L %s -u %s %s %s' % (cuffdir+'/cuffdiff', cfg.genome, condition.keys()[0]+','+condition.keys()[1], cuffdir+'/merged_asm/merged.gtf', condition.values()[0], condition.values()[1])
NGSTools.writeCommands(command, cuffdir+'/cuffdiff.sh', _run)
if DESeq2:
# deseq2 #
deseqDir = os.path.join(args.outDir, 'DESeq')
try:
os.mkdir(deseqDir)
except:
def processSample(sampleName, sampleNameDict):
'''pipeline for sample '''
# process communication
mng = Manager()
libraryBamFileList = mng.list()
# This libraryBamFileList is a list contains seval bams from one sample.
# init mult-processer
record = []
for sampleID in sampleNameDict:
Processer = Process(name=sampleID,
target=processSampleFromLibarary,
args=(
sampleID,
sampleNameDict[sampleID],
libraryBamFileList,
))
Processer.start()
record.append(Processer)
# wait for processer
for proc in record:
proc.join()
###################### 2.1 post mapping ####################
bamFileDir = os.path.dirname(libraryBamFileList[0])
mergedBamFilePath = os.path.join(bamFileDir, "%s_merged.bam" % sampleName)
finalBamFilePath = mergedBamFilePath
# merge the bam from each lane to one final bam
command = NGSTools.picard_merge(libraryBamFileList, mergedBamFilePath, cfg)
NGSTools.writeCommands(command,
bamFileDir + '/picard_mergebam_' + sampleName +
'.sh',
run=_run)
###################### 2.2. filter bam ######################
#command = 'samtools view -Sb -h -f 2 -q 10 %s > %s ' % (mergedBamFilePath, finalBamFilePath)
#command = 'samtools view -Sb -h -q 10 %s > %s ' % (mergedBamFilePath, finalBamFilePath)
#NGSTools.writeCommands(command, bamFileDir+'/filterBam_'+sampleName+'.sh', run=_run)
###################### 3. romove duplicates ##################
if RMDUP:
command = NGSTools.picard_rmdup(mergedBamFilePath, True, cfg)
NGSTools.writeCommands(command,
bamFileDir + '/picard_rmdup_' + sampleName +
'.sh',
run=_run)
finalBamFilePath = re.sub(r'.bam$', '.rmdup.bam', finalBamFilePath)
####################### 4. call SNP ######################
if MPILEUP:
SNP_out = os.path.join(args.outDir, "SNP", sampleName)
NGSTools._mkdir(SNP_out)
command, rawVcf = NGSTools.bcftools_call(finalBamFilePath,
cfg,
outdir=SNP_out,
sampleName=sampleName)
NGSTools.writeCommands(command,
SNP_out + '/bcftools_call_' + sampleName +
'.sh',
run=_run)
outVcf = rawVcf.replace("vcf$", "flt.vcf")
command = NGSTools.bcftools_filter(rawVcf, outVcf, cfg)
NGSTools.writeCommands(command,
SNP_out + '/bcftools_filter_' + sampleName +
'.sh',
run=_run)
