def sambamba(inbams,
outbam,
tech='docker',
input_parameters={},
remove_inbams=False):
for param_i in DEFAULT_PARAMS:
if param_i not in input_parameters:
input_parameters[param_i] = DEFAULT_PARAMS[param_i]
logdir = os.path.join(input_parameters['output_directory'], 'logs')
outfile = os.path.join(logdir, input_parameters['script'])
all_paths = list(inbams) + [
outbam,
]
merge_line, fileDict = container.container_params(
input_parameters['sambamba_image'],
tech=tech,
files=all_paths,
extra_args=input_parameters['extra_docker_options'])
mounted_outbam = fileDict[outbam]['mount_path']
infile_string = ' '.join(
[fileDict[file_i]['mount_path'] for file_i in inbams])
with open(outfile, 'w') as out:
out.write("#!/bin/bash\n\n")
out.write(f'#$ -o {logdir}\n')
out.write(f'#$ -e {logdir}\n')
out.write('#$ -S /bin/bash\n')
out.write('#$ -l h_vmem={}G\n'.format(input_parameters['MEM']))
out.write('set -e\n\n')
out.write(
'echo -e "Start at `date +"%Y/%m/%d %H:%M:%S"`" 1>&2\n\n'
) # Do not change this: picard_fractional uses this to end the copying.
out.write(f'{merge_line} \\\n')
out.write('sambamba merge -t {} {} {}\n\n'.format(
input_parameters['threads'], mounted_outbam, infile_string))
if remove_inbams:
out.write('rm {}\n\n'.format(' '.join(inbams)))
out.write('\necho -e "Done at `date +"%Y/%m/%d %H:%M:%S"`" 1>&2\n')
# "Run" the script that was generated
command_line = '{} {}'.format(input_parameters['action'], outfile)
returnCode = subprocess.call(command_line, shell=True)
return outfile
def run_SomaticSeq(input_parameters, tech='docker'):
DEFAULT_PARAMS = {'MEM': '4G', 'inclusion_region': None, 'exclusion_region': None, 'output_directory' : os.curdir, 'somaticseq_directory': 'SomaticSeq', 'action': 'echo', 'dbsnp' : None, 'cosmic': None, 'snv_classifier': None, 'indel_classifier': None, 'truth_snv': None, 'truth_indel': None, 'somaticseq_arguments': '', 'train_somaticseq': False, 'somaticseq_algorithm': 'xgboost'}
for param_i in DEFAULT_PARAMS:
if param_i not in input_parameters:
input_parameters[param_i] = DEFAULT_PARAMS[param_i]
all_paths = []
for path_i in input_parameters['bam'], input_parameters['genome_reference'], input_parameters['output_directory'], input_parameters['inclusion_region'], input_parameters['exclusion_region'], input_parameters['dbsnp'], input_parameters['cosmic'], input_parameters['snv_classifier'], input_parameters['indel_classifier'], input_parameters['truth_snv'], input_parameters['truth_indel']:
if path_i:
all_paths.append( path_i )
container_line, fileDict = container.container_params( f'lethalfang/somaticseq:{VERSION}', tech=tech, files=all_paths, extra_args=input_parameters['extra_docker_options'] )
# Mounted paths for all the input files and output directory:
mounted_genome_reference = fileDict[ input_parameters['genome_reference'] ]['mount_path']
mounted_tumor_bam = fileDict[ input_parameters['bam'] ]['mount_path']
mounted_outdir = fileDict[ input_parameters['output_directory'] ]['mount_path']
outdir = os.path.join(input_parameters['output_directory'], input_parameters['somaticseq_directory'])
logdir = os.path.join(outdir, 'logs')
outfile = os.path.join(logdir, input_parameters['script'] )
mutect2 = '{}/MuTect2.vcf'.format(mounted_outdir)
varscan = '{}/VarScan2.vcf'.format(mounted_outdir)
vardict = '{}/VarDict.vcf'.format(mounted_outdir)
lofreq = '{}/LoFreq.vcf'.format(mounted_outdir)
scalpel = '{}/Scalpel.vcf'.format(mounted_outdir)
strelka = '{}/Strelka/results/variants/variants.vcf.gz'.format(mounted_outdir)
os.makedirs(logdir, exist_ok=True)
with open(outfile, 'w') as out:
out.write( "#!/bin/bash\n\n" )
out.write(f'#$ -o {logdir}\n' )
out.write(f'#$ -e {logdir}\n' )
out.write( '#$ -S /bin/bash\n' )
out.write( '#$ -l h_vmem={}\n'.format( input_parameters['MEM'] ) )
out.write( 'set -e\n\n' )
out.write( 'echo -e "Start at `date +"%Y/%m/%d %H:%M:%S"`" 1>&2\n\n' )
#out.write( 'docker pull lethalfang/somaticseq:{VERSION} \n\n'.format(VERSION=VERSION) )
out.write(f'{container_line} \\\n' )
out.write( '/opt/somaticseq/somaticseq/run_somaticseq.py \\\n' )
if input_parameters['train_somaticseq'] and input_parameters['threads'] == 1:
out.write( '--somaticseq-train --algorithm {} \\\n'.format(input_parameters['somaticseq_algorithm']) )
out.write( '--output-directory {} \\\n'.format( os.path.join(mounted_outdir, input_parameters['somaticseq_directory']) ) )
out.write( '--genome-reference {} \\\n'.format(mounted_genome_reference) )
if input_parameters['inclusion_region']:
mounted_inclusion = fileDict[ input_parameters['inclusion_region'] ]['mount_path']
out.write( '--inclusion-region {} \\\n'.format(mounted_inclusion) )
if input_parameters['exclusion_region']:
mounted_exclusion = fileDict[ input_parameters['exclusion_region'] ]['mount_path']
out.write( '--exclusion-region {} \\\n'.format(input_parameters['exclusion_region']) )
if input_parameters['cosmic']:
mounted_cosmic = fileDict[ input_parameters['cosmic'] ]['mount_path']
out.write( '--cosmic-vcf {} \\\n'.format(mounted_cosmic) )
if input_parameters['dbsnp']:
mounted_dbsnp = fileDict[ input_parameters['dbsnp'] ]['mount_path']
out.write( '--dbsnp-vcf {} \\\n'.format(input_parameters['dbsnp_vcf']) )
if input_parameters['snv_classifier'] or input_parameters['indel_classifier']:
out.write( '--algorithm {} \\\n'.format(input_parameters['somaticseq_algorithm']) )
if input_parameters['snv_classifier']:
out.write( '--classifier-snv {} \\\n'.format( fileDict[ input_parameters['snv_classifier'] ]['mount_path'] ) )
if input_parameters['indel_classifier']:
out.write( '--classifier-indel {} \\\n'.format( fileDict[ input_parameters['indel_classifier'] ]['mount_path'] ) )
if input_parameters['truth_snv']:
out.write( '--truth-snv {} \\\n'.format( fileDict[ input_parameters['truth_snv'] ]['mount_path'] ) )
if input_parameters['truth_indel']:
out.write( '--truth-indel {} \\\n'.format( fileDict[ input_parameters['truth_indel'] ]['mount_path'] ) )
if input_parameters['somaticseq_algorithm']:
out.write( '--algorithm {} \\\n'.format(input_parameters['somaticseq_algorithm']) )
if input_parameters['somaticseq_arguments']:
out.write( '{} \\\n'.format(input_parameters['somaticseq_arguments']) )
out.write( 'single \\\n' )
out.write( '--bam-file {} \\\n'.format(mounted_tumor_bam) )
if input_parameters['run_mutect2']:
out.write( '--mutect2-vcf {} \\\n'.format(mutect2) )
if input_parameters['run_varscan2']:
out.write( '--varscan-vcf {} \\\n'.format(varscan) )
if input_parameters['run_vardict']:
out.write( '--vardict-vcf {} \\\n'.format(vardict) )
if input_parameters['run_lofreq']:
out.write( '--lofreq-vcf {} \\\n'.format(lofreq) )
if input_parameters['run_scalpel']:
out.write( '--scalpel-vcf {} \\\n'.format(scalpel) )
if input_parameters['run_strelka2']:
out.write( '--strelka-vcf {} \\\n'.format(strelka) )
out.write( '\necho -e "Done at `date +"%Y/%m/%d %H:%M:%S"`" 1>&2\n' )
# "Run" the script that was generated
command_line = '{} {}'.format( input_parameters['action'], outfile )
returnCode = subprocess.call( command_line, shell=True )
return outfile
def merge_results(input_parameters, tech='docker'):
DEFAULT_PARAMS = {'MEM': '4G', 'output_directory': os.curdir, 'somaticseq_directory': 'SomaticSeq', 'action': 'echo', 'script': 'mergeResults.{}.cmd'.format(ts), 'snv_classifier': None, 'indel_classifier': None, 'truth_snv': None, 'truth_indel': None, 'somaticseq_arguments': '', 'train_somaticseq': False, 'somaticseq_algorithm': 'xgboost'}
for param_i in DEFAULT_PARAMS:
if param_i not in input_parameters:
input_parameters[param_i] = DEFAULT_PARAMS[param_i]
all_paths = []
for path_i in input_parameters['genome_reference'], input_parameters['output_directory'], input_parameters['snv_classifier'], input_parameters['indel_classifier'], input_parameters['truth_snv'], input_parameters['truth_indel']:
if path_i:
all_paths.append( path_i )
container_line, fileDict = container.container_params( f'lethalfang/somaticseq:{VERSION}', tech=tech, files=all_paths, extra_args=input_parameters['extra_docker_options'] )
# Mounted paths for all the input files and output directory:
mounted_outdir = fileDict[ input_parameters['output_directory'] ]['mount_path']
prjdir = input_parameters['output_directory']
logdir = os.path.join(prjdir, 'logs')
outfile = os.path.join(logdir, input_parameters['script'] )
mutect2 = mounted_outdir + '/{}/MuTect2.vcf'
varscan = mounted_outdir + '/{}/VarScan2.vcf'
vardict = mounted_outdir + '/{}/VarDict.vcf'
lofreq = mounted_outdir + '/{}/LoFreq.vcf'
scalpel = mounted_outdir + '/{}/Scalpel.vcf'
strelka = mounted_outdir + '/{}/Strelka/results/variants/variants.vcf.gz'
somaticdir = input_parameters['somaticseq_directory']
os.makedirs(logdir, exist_ok=True)
with open(outfile, 'w') as out:
out.write( "#!/bin/bash\n\n" )
out.write(f'#$ -o {logdir}\n' )
out.write(f'#$ -e {logdir}\n' )
out.write( '#$ -S /bin/bash\n' )
out.write( '#$ -l h_vmem={}\n'.format( input_parameters['MEM'] ) )
out.write( 'set -e\n\n' )
out.write( 'echo -e "Start at `date +"%Y/%m/%d %H:%M:%S"`" 1>&2\n\n' )
if input_parameters['run_mutect2']:
out.write(f'{container_line} \\\n' )
out.write( 'concat.py --bgzip-output -infiles \\\n' )
for i in range(1, input_parameters['threads']+1):
out.write( mutect2.format(i) + ' ' )
out.write( '\\\n' )
out.write('-outfile {}/MuTect2.vcf\n\n'.format(mounted_outdir) )
if input_parameters['run_varscan2']:
out.write(f'{container_line} \\\n' )
out.write( 'concat.py --bgzip-output -infiles \\\n' )
for i in range(1, input_parameters['threads']+1):
out.write( varscan.format(i) + ' ' )
out.write( '\\\n' )
out.write('-outfile {}/VarScan2.vcf\n\n'.format(mounted_outdir) )
if input_parameters['run_vardict']:
out.write(f'{container_line} \\\n' )
out.write( 'concat.py --bgzip-output -infiles \\\n' )
for i in range(1, input_parameters['threads']+1):
out.write( vardict.format(i) + ' ' )
out.write( '\\\n' )
out.write('-outfile {}/VarDict.vcf\n\n'.format(mounted_outdir) )
if input_parameters['run_lofreq']:
out.write(f'{container_line} \\\n' )
out.write( 'concat.py --bgzip-output -infiles \\\n' )
for i in range(1, input_parameters['threads']+1):
out.write( lofreq.format(i) + ' ' )
out.write( '\\\n' )
out.write('-outfile {}/LoFreq.vcf\n\n'.format(mounted_outdir) )
if input_parameters['run_scalpel']:
out.write(f'{container_line} \\\n' )
out.write( 'concat.py --bgzip-output -infiles \\\n' )
for i in range(1, input_parameters['threads']+1):
out.write( scalpel.format(i) + ' ' )
out.write( '\\\n' )
out.write('-outfile {}/Scalpel.vcf\n\n'.format(mounted_outdir) )
if input_parameters['run_strelka2']:
out.write(f'{container_line} \\\n' )
out.write( 'concat.py --bgzip-output -infiles \\\n' )
for i in range(1, input_parameters['threads']+1):
out.write( strelka.format(i) + ' ' )
out.write( '\\\n' )
out.write('-outfile {}/Strelka.vcf\n\n'.format(mounted_outdir) )
###### SomaticSeq #####
if input_parameters['run_somaticseq']:
# Ensemble.sSNV.tsv
out.write(f'{container_line} \\\n' )
out.write( 'concat.py -infiles \\\n' )
for i in range(1, input_parameters['threads']+1):
out.write( '{}/{}/{}/Ensemble.sSNV.tsv'.format(mounted_outdir, i, somaticdir) + ' ' )
out.write( '\\\n' )
out.write('-outfile {}/Ensemble.sSNV.tsv\n\n'.format(mounted_outdir) )
# Ensemble.sINDEL.tsv
out.write(f'{container_line} \\\n' )
out.write( 'concat.py -infiles \\\n' )
for i in range(1, input_parameters['threads']+1):
out.write( '{}/{}/{}/Ensemble.sINDEL.tsv'.format(mounted_outdir, i, somaticdir) + ' ' )
out.write( '\\\n' )
out.write('-outfile {}/Ensemble.sINDEL.tsv\n\n'.format(mounted_outdir) )
# If asked to create classifier, do it here when TSV files are combined
if input_parameters['train_somaticseq'] and input_parameters['truth_snv']:
out.write(f'{container_line} \\\n' )
if input_parameters['somaticseq_algorithm'] == 'ada':
out.write( 'ada_model_builder_ntChange.R {}/Ensemble.sSNV.tsv\n\n'.format( mounted_outdir) )
else:
out.write( 'somatic_xgboost.py train -threads {} -tsvs {}/Ensemble.sSNV.tsv\n\n'.format(input_parameters['threads'], mounted_outdir) )
if input_parameters['train_somaticseq'] and input_parameters['truth_indel']:
out.write(f'{container_line} \\\n' )
if input_parameters['somaticseq_algorithm'] == 'ada':
out.write( 'ada_model_builder_ntChange.R {}/Ensemble.sINDEL.tsv\n\n'.format( mounted_outdir) )
else:
out.write( 'somatic_xgboost.py train -threads {} -tsvs {}/Ensemble.sINDEL.tsv\n\n'.format(input_parameters['threads'], mounted_outdir) )
# If in prediction mode, combine SSeq.Classified.sSNV.vcf, else Consensus.sSNV.vcf
if input_parameters['snv_classifier']:
out.write(f'{container_line} \\\n' )
out.write( 'concat.py --bgzip-output -infiles \\\n' )
for i in range(1, input_parameters['threads']+1):
out.write( '{}/{}/{}/SSeq.Classified.sSNV.vcf'.format(mounted_outdir, i, somaticdir) + ' ' )
out.write( '\\\n' )
out.write('-outfile {}/SSeq.Classified.sSNV.vcf\n\n'.format(mounted_outdir) )
# SSeq.Classified.sSNV.tsv
out.write(f'{container_line} \\\n' )
out.write( 'concat.py --bgzip-output -infiles \\\n' )
for i in range(1, input_parameters['threads']+1):
out.write( '{}/{}/{}/SSeq.Classified.sSNV.tsv'.format(mounted_outdir, i, somaticdir) + ' ' )
out.write( '\\\n' )
out.write('-outfile {}/SSeq.Classified.sSNV.tsv\n\n'.format(mounted_outdir) )
# Consensus mode: Consensus.sSNV.vcf
else:
out.write(f'{container_line} \\\n' )
out.write( 'concat.py --bgzip-output -infiles \\\n' )
for i in range(1, input_parameters['threads']+1):
out.write( '{}/{}/{}/Consensus.sSNV.vcf'.format(mounted_outdir, i, somaticdir) + ' ' )
out.write( '\\\n' )
out.write('-outfile {}/Consensus.sSNV.vcf\n\n'.format(mounted_outdir) )
# If in prediction mode, combine SSeq.Classified.sINDEL.vcf, else Consensus.sINDEL.vcf
if input_parameters['indel_classifier']:
out.write(f'{container_line} \\\n' )
out.write( 'concat.py --bgzip-output -infiles \\\n' )
for i in range(1, input_parameters['threads']+1):
out.write( '{}/{}/{}/SSeq.Classified.sINDEL.vcf'.format(mounted_outdir, i, somaticdir) + ' ' )
out.write( '\\\n' )
out.write('-outfile {}/SSeq.Classified.sINDEL.vcf\n\n'.format(mounted_outdir) )
# SSeq.Classified.sINDEL.tsv
out.write(f'{container_line} \\\n' )
out.write( 'concat.py --bgzip-output -infiles \\\n' )
for i in range(1, input_parameters['threads']+1):
out.write( '{}/{}/{}/SSeq.Classified.sINDEL.tsv'.format(mounted_outdir, i, somaticdir) + ' ' )
out.write( '\\\n' )
out.write('-outfile {}/SSeq.Classified.sINDEL.tsv\n\n'.format(mounted_outdir) )
# Consensus mode: Consensus.sINDEL.vcf
else:
out.write(f'{container_line} \\\n' )
out.write( 'concat.py --bgzip-output -infiles \\\n' )
for i in range(1, input_parameters['threads']+1):
out.write( '{}/{}/{}/Consensus.sINDEL.vcf'.format(mounted_outdir, i, somaticdir) + ' ' )
out.write( '\\\n' )
out.write('-outfile {}/Consensus.sINDEL.vcf\n\n'.format(mounted_outdir) )
out.write( '\necho -e "Done at `date +"%Y/%m/%d %H:%M:%S"`" 1>&2\n' )
command_line = '{} {}'.format( input_parameters['action'], outfile )
returnCode = subprocess.call( command_line, shell=True )
return outfile
def bwa(input_parameters, tech='docker'):
if input_parameters['in_fastq2']:
paired_end = True
else:
paired_end = False
for param_i in DEFAULT_PARAMS:
if param_i not in input_parameters:
input_parameters[param_i] = DEFAULT_PARAMS[param_i]
#
logdir = os.path.join(input_parameters['output_directory'], 'logs')
outfile = os.path.join(logdir, input_parameters['script'])
all_paths = []
for path_i in input_parameters['output_directory'], input_parameters[
'genome_reference'], input_parameters[
'in_fastq1'], input_parameters['in_fastq2']:
if path_i:
all_paths.append(path_i)
bwa_line, fileDict = container.container_params(
input_parameters['bwa_image'],
tech=tech,
files=all_paths,
extra_args=input_parameters['extra_docker_options'])
# Mounted paths for all the input files and output directory:
mounted_outdir = fileDict[
input_parameters['output_directory']]['mount_path']
mounted_reference = fileDict[
input_parameters['genome_reference']]['mount_path']
mounted_fq1 = fileDict[input_parameters['in_fastq1']]['mount_path']
mounted_fq2 = fileDict[input_parameters['in_fastq2']]['mount_path']
temporary_files = []
with open(outfile, 'w') as out:
out.write("#!/bin/bash\n\n")
out.write(f'#$ -o {logdir}\n')
out.write(f'#$ -e {logdir}\n')
out.write('#$ -S /bin/bash\n')
out.write('#$ -l h_vmem={}G\n'.format(input_parameters['MEM'] *
input_parameters['threads']))
out.write('set -e\n\n')
out.write('echo -e "Start at `date +"%Y/%m/%d %H:%M:%S"`" 1>&2\n\n')
out.write(f'{bwa_line} bash -c \\\n')
out.write('"bwa mem \\\n')
out.write('-R \'{}\' \\\n'.format(input_parameters['bam_header']))
out.write('-M {} -t {} \\\n'.format(
input_parameters['extra_bwa_arguments'],
input_parameters['threads']))
out.write('{} \\\n'.format(mounted_reference))
out.write('{} \\\n'.format(mounted_fq1))
if paired_end:
out.write('{} \\\n'.format(mounted_fq2))
out.write('| samtools view -Sbh - \\\n')
out.write(
'| samtools sort -m {MEM}G --threads {THREADS} -o {DIR}/{OUTFILE}"\n\n'
.format(MEM=math.ceil(input_parameters['MEM'] / 2),
THREADS=math.ceil(input_parameters['threads'] / 2),
DIR=mounted_outdir,
OUTFILE=input_parameters['out_bam']))
out.write(f'{bwa_line} \\\n')
out.write('samtools index -@{} {}\n'.format(
input_parameters['threads'],
os.path.join(mounted_outdir, input_parameters['out_bam'])))
out.write('\necho -e "Done at `date +"%Y/%m/%d %H:%M:%S"`" 1>&2\n')
# "Run" the script that was generated
command_line = '{} {}'.format(input_parameters['action'], outfile)
returnCode = subprocess.call(command_line, shell=True)
return outfile
def tumor_normal(input_parameters=DEFAULT_PARAMS, tech='docker'):
for param_i in DEFAULT_PARAMS:
if param_i not in input_parameters:
input_parameters[param_i] = DEFAULT_PARAMS[param_i]
# The following are required:
assert os.path.exists(input_parameters['normal_bam'])
assert os.path.exists(input_parameters['tumor_bam'])
assert os.path.exists(input_parameters['genome_reference'])
logdir = os.path.join(input_parameters['output_directory'], 'logs')
outfile = os.path.join(logdir, input_parameters['script'])
all_paths = []
for path_i in input_parameters['normal_bam'], input_parameters[
'tumor_bam'], input_parameters[
'genome_reference'], input_parameters[
'output_directory'], input_parameters['inclusion_region']:
if path_i:
all_paths.append(path_i)
container_line, fileDict = container.container_params(
input_parameters['strelka2_image'],
tech=tech,
files=all_paths,
extra_args=input_parameters['extra_docker_options'])
# Mounted paths for all the input files and output directory:
mounted_genome_reference = fileDict[
input_parameters['genome_reference']]['mount_path']
mounted_tumor_bam = fileDict[input_parameters['tumor_bam']]['mount_path']
mounted_normal_bam = fileDict[input_parameters['normal_bam']]['mount_path']
mounted_outdir = fileDict[
input_parameters['output_directory']]['mount_path']
if input_parameters['inclusion_region']:
mounted_inclusion = fileDict[
input_parameters['inclusion_region']]['mount_path']
bed_gz = fileDict[
input_parameters['inclusion_region']]['filename'] + '.gz'
with open(outfile, 'w') as out:
out.write("#!/bin/bash\n\n")
out.write(f'#$ -o {logdir}\n')
out.write(f'#$ -e {logdir}\n')
out.write('#$ -S /bin/bash\n')
out.write('#$ -l h_vmem={}\n'.format(input_parameters['MEM']))
out.write('set -e\n\n')
out.write('echo -e "Start at `date +"%Y/%m/%d %H:%M:%S"`" 1>&2\n\n')
# Make .bed.gz out of .bed files using tabix:
tabix_line, tabixDict = container.container_params(
'lethalfang/tabix:1.7', tech, all_paths)
tabix_selector = tabixDict[
input_parameters['inclusion_region']]['mount_path']
tabix_outdir = tabixDict[
input_parameters['output_directory']]['mount_path']
out.write(
'{DOCKER_LINE} bash -c "cat {SELECTOR} | bgzip > {OUTDIR}/{BEDGZ}\"\n'
.format(DOCKER_LINE=tabix_line,
SELECTOR=tabix_selector,
OUTDIR=tabix_outdir,
BEDGZ=bed_gz))
out.write('{DOCKER_LINE} tabix -f {OUTDIR}/{BEDGZ}\n\n'.format(
DOCKER_LINE=tabix_line, OUTDIR=tabix_outdir, BEDGZ=bed_gz))
out.write(f'{container_line} \\\n')
out.write('/opt/strelka/bin/configureStrelkaSomaticWorkflow.py \\\n')
out.write('--tumorBam={} \\\n'.format(mounted_tumor_bam))
out.write('--normalBam={} \\\n'.format(mounted_normal_bam))
out.write('--referenceFasta={} \\\n'.format(mounted_genome_reference))
out.write('--callMemMb={} \\\n'.format(
eval(input_parameters['MEM'].rstrip('G')) * 1024))
out.write('--callRegions={}/{} \\\n'.format(mounted_outdir, bed_gz))
if input_parameters['exome']:
out.write('--exome \\\n')
if input_parameters['strelka_config_arguments']:
out.write('{} \\\n'.format(
input_parameters['strelka_config_arguments']))
out.write('--runDir={}/{}\n\n'.format(mounted_outdir,
input_parameters['outdir_name']))
out.write(f'{container_line} \\\n')
out.write('{}/{}/runWorkflow.py -m local -j 1 {}\n'.format(
mounted_outdir, input_parameters['outdir_name'],
input_parameters['strelka_run_arguments']))
out.write('\necho -e "Done at `date +"%Y/%m/%d %H:%M:%S"`" 1>&2\n')
# "Run" the script that was generated
command_line = '{} {}'.format(input_parameters['action'], outfile)
returnCode = subprocess.call(command_line, shell=True)
return outfile
def alienTrimmer(input_parameters, tech='docker'):
if input_parameters['in_fastq2']:
paired_end = True
else:
paired_end = False
for param_i in DEFAULT_PARAMS:
if param_i not in input_parameters:
input_parameters[param_i] = DEFAULT_PARAMS[param_i]
#
logdir = os.path.join(input_parameters['output_directory'], 'logs')
outfile = os.path.join(logdir, input_parameters['script'])
all_paths = []
for path_i in input_parameters['output_directory'], input_parameters[
'in_fastq1'], input_parameters['in_fastq2']:
if path_i:
all_paths.append(path_i)
trim_line, fileDict = container.container_params(
input_parameters['alienTrimmerImage'],
tech=tech,
files=all_paths,
extra_args=input_parameters['extra_docker_options'])
# Mounted paths for all the input files and output directory:
mounted_outdir = fileDict[
input_parameters['output_directory']]['mount_path']
temporary_files = []
with open(outfile, 'w') as out:
out.write("#!/bin/bash\n\n")
out.write(f'#$ -o {logdir}\n')
out.write(f'#$ -e {logdir}\n')
out.write('#$ -S /bin/bash\n')
out.write('#$ -l h_vmem={}G\n'.format(input_parameters['MEM']))
out.write('set -e\n\n')
out.write('echo -e "Start at `date +"%Y/%m/%d %H:%M:%S"`" 1>&2\n\n')
# AlienTrimmer does not do bgzipped fastq files, unfortunately:
if input_parameters['in_fastq1'].endswith('.gz'):
out_fastq_1 = uuid.uuid4().hex + '.fastq'
out_fastq_2 = uuid.uuid4().hex + '.fastq'
if paired_end:
tabix_line, tabixDict = container.container_params(
'lethalfang/tabix:1.7', tech,
(input_parameters['output_directory'],
input_parameters['in_fastq1'],
input_parameters['in_fastq2']))
else:
tabix_line, tabixDict = container.container_params(
'lethalfang/tabix:1.7', tech,
(input_parameters['output_directory'],
input_parameters['in_fastq1']))
tabix_outdir = tabixDict[
input_parameters['output_directory']]['mount_path']
tabix_fq1 = tabixDict[input_parameters['in_fastq1']]['mount_path']
out.write(f'{tabix_line} bash -c \\\n')
out.write('"gunzip -c {} > {}/{}"\n\n'.format(
tabix_fq1, tabix_outdir, out_fastq_1))
mounted_fq1 = os.path.join(mounted_outdir, out_fastq_1)
temporary_files.append(out_fastq_1)
if paired_end:
tabix_fq2 = tabixDict[
input_parameters['in_fastq2']]['mount_path']
out.write(f'{tabix_line} bash -c \\\n')
out.write('"gunzip -c {} > {}/{}"\n\n'.format(
tabix_fq2, tabix_outdir, out_fastq_2))
mounted_fq2 = os.path.join(mounted_outdir, out_fastq_2)
temporary_files.append(out_fastq_2)
else:
mounted_fq1 = fileDict[input_parameters['in_fastq1']]['mount_path']
if paired_end:
mounted_fq2 = fileDict[
input_parameters['in_fastq2']]['mount_path']
out.write(f'{trim_line} \\\n')
out.write('/opt/AlienTrimmer_0.4.0/src/AlienTrimmer \\\n')
if paired_end:
trimmed_fq1 = uuid.uuid4().hex + '.fastq'
trimmed_fq2 = uuid.uuid4().hex + '.fastq'
singleton = uuid.uuid4().hex + '.fastq'
out.write('-if {} -ir {} \\\n'.format(mounted_fq1, mounted_fq2))
out.write('-of {}/{} -or {}/{} \\\n'.format(
mounted_outdir, trimmed_fq1, mounted_outdir, trimmed_fq2))
out.write('-os {}/{} \\\n'.format(mounted_outdir, singleton))
temporary_files.extend([trimmed_fq1, trimmed_fq2, singleton])
else:
trimmed_fq1 = uuid.uuid4().hex + '.fastq'
out.write('-i {} \\\n'.write(mounted_fq1))
out.write('-o {}/{} \\\n'.write(mounted_outdir, trimmed_fq1))
temporary_files.append(trimmed_fq1)
out.write('-c {} \\\n'.format(input_parameters['adapter']))
out.write('-l {}\n\n'.format(input_parameters['minimum_length']))
out.write(f'{tabix_line} bash -c \\\n')
out.write('"cat {}/{} | bgzip -@{} > {}/{}"\n'.format(
tabix_outdir, trimmed_fq1, input_parameters['threads'],
tabix_outdir, input_parameters['out_fastq1_name']))
if paired_end:
out.write(f'{tabix_line} bash -c \\\n')
out.write('"cat {}/{} | bgzip -@{} > {}/{}"\n'.format(
tabix_outdir, trimmed_fq2, input_parameters['threads'],
tabix_outdir, input_parameters['out_fastq2_name']))
out.write(f'{tabix_line} bash -c \\\n')
out.write('"cat {}/{} | bgzip -@{} > {}/{}"\n'.format(
tabix_outdir, singleton, input_parameters['threads'],
tabix_outdir, input_parameters['out_singleton_name']))
out.write('\n')
for file_i in temporary_files:
out.write('rm {}\n'.format(
os.path.join(input_parameters['output_directory'], file_i)))
# Remove untrimmed files:
if input_parameters['remove_untrimmed']:
out.write('\n')
out.write('rm {}\n'.format(input_parameters['in_fastq1']))
if input_parameters['in_fastq2']:
out.write('rm {}\n'.format(input_parameters['in_fastq2']))
out.write('\necho -e "Done at `date +"%Y/%m/%d %H:%M:%S"`" 1>&2\n')
# "Run" the script that was generated
command_line = '{} {}'.format(input_parameters['action'], outfile)
returnCode = subprocess.call(command_line, shell=True)
return outfile
def tumor_normal(input_parameters, tech='docker'):
for param_i in DEFAULT_PARAMS:
if param_i not in input_parameters:
input_parameters[param_i] = DEFAULT_PARAMS[param_i]
# The following are required:
assert os.path.exists(input_parameters['normal_bam'])
assert os.path.exists(input_parameters['tumor_bam'])
assert os.path.exists(input_parameters['genome_reference'])
logdir = os.path.join(input_parameters['output_directory'], 'logs')
outfile = os.path.join(logdir, input_parameters['script'])
all_paths = []
for path_i in input_parameters['normal_bam'], input_parameters[
'tumor_bam'], input_parameters[
'genome_reference'], input_parameters[
'output_directory'], input_parameters['inclusion_region']:
if path_i:
all_paths.append(path_i)
container_line, fileDict = container.container_params(
input_parameters['vardict_image'],
tech=tech,
files=all_paths,
extra_args=input_parameters['extra_docker_options'])
minVAF = input_parameters['minimum_VAF']
total_bases = 0
num_lines = 0
if input_parameters['inclusion_region']:
bed_file = input_parameters['inclusion_region']
with open(bed_file) as bed:
line_i = bed.readline().rstrip()
while line_i.startswith('track'):
line_i = bed.readline().rstrip()
while line_i:
item = line_i.rstrip().split('\t')
total_bases = total_bases + int(item[2]) - int(item[1])
num_lines += 1
line_i = bed.readline().rstrip()
else:
fai_file = input_parameters['genome_reference'] + '.fai'
bed_file = os.path.join(input_parameters['output_directory'],
'genome.bed')
with open(fai_file) as fai, open(bed_file, 'w') as wgs_bed:
for line_i in fai:
item = line_i.split('\t')
total_bases += int(item[1])
num_lines += 1
wgs_bed.write('{}\t{}\t{}\n'.format(item[0], '0', item[1]))
# However the "bed_file" is defined here, create a dockered line and mount dictionary for it:
bed_split_line, bedDict = container.container_params(
'lethalfang/somaticseq:{}'.format(VERSION), tech,
(bed_file, input_parameters['output_directory']))
# Mounted paths for all the input files and output directory:
mounted_genome_reference = fileDict[
input_parameters['genome_reference']]['mount_path']
mounted_tumor_bam = fileDict[input_parameters['tumor_bam']]['mount_path']
mounted_normal_bam = fileDict[input_parameters['normal_bam']]['mount_path']
mounted_outdir = fileDict[
input_parameters['output_directory']]['mount_path']
mounted_bed = bedDict[bed_file]['mount_path']
with open(outfile, 'w') as out:
out.write("#!/bin/bash\n\n")
out.write(f'#$ -o {logdir}\n')
out.write(f'#$ -e {logdir}\n')
out.write('#$ -S /bin/bash\n')
out.write('#$ -l h_vmem={}\n'.format(input_parameters['MEM']))
out.write('set -e\n\n')
out.write('echo -e "Start at `date +"%Y/%m/%d %H:%M:%S"`" 1>&2\n\n')
# Decide if Bed file needs to be "split" such that each line has a small enough region
if input_parameters['process_bed'] or total_bases / num_lines > 50000:
out.write(f'{bed_split_line} \\\n')
out.write(
'/opt/somaticseq/somaticseq/utilities/split_mergedBed.py \\\n')
out.write('-infile {} -outfile {}/split_regions.bed\n\n'.format(
mounted_bed,
bedDict[input_parameters['output_directory']]['mount_path']))
bed_file = '{}/split_regions.bed'.format(mounted_outdir)
out.write(f'{container_line} bash -c \\\n')
out.write('"/opt/VarDict-1.7.0/bin/VarDict \\\n')
if input_parameters['vardict_arguments']:
out.write('{} \\\n'.format(input_parameters['vardict_arguments']))
out.write('-G {} \\\n'.format(mounted_genome_reference))
out.write('-f {} -h \\\n'.format(minVAF))
out.write('-b \'{}|{}\' \\\n'.format(mounted_tumor_bam,
mounted_normal_bam))
out.write('-Q 1 -c 1 -S 2 -E 3 -g 4 {} \\\n'.format(bed_file))
out.write('> {}/vardict.var"\n\n'.format(mounted_outdir))
out.write('\n')
out.write(f'{container_line} \\\n')
out.write(
'bash -c "cat {}/vardict.var | awk \'NR!=1\' | /opt/VarDict/testsomatic.R | /opt/VarDict/var2vcf_paired.pl -N \'TUMOR|NORMAL\' -f {} \\\n'
.format(mounted_outdir, minVAF))
out.write('> {}/{}"\n\n'.format(mounted_outdir,
input_parameters['outfile']))
out.write('\necho -e "Done at `date +"%Y/%m/%d %H:%M:%S"`" 1>&2\n')
# "Run" the script that was generated
command_line = '{} {}'.format(input_parameters['action'], outfile)
returnCode = subprocess.call(command_line, shell=True)
return outfile
def tumor_normal(input_parameters=DEFAULT_PARAMS, tech='docker'):
for param_i in DEFAULT_PARAMS:
if param_i not in input_parameters:
input_parameters[param_i] = DEFAULT_PARAMS[param_i]
# The following are required:
assert os.path.exists(input_parameters['tumor_bam'])
assert os.path.exists(input_parameters['normal_bam'])
assert os.path.exists(input_parameters['genome_reference'])
assert os.path.exists(input_parameters['dbsnp_gz'])
assert os.path.exists(input_parameters['dbsnp_gz'] + '.tbi')
logdir = os.path.join(input_parameters['output_directory'], 'logs')
outfile = os.path.join(logdir, input_parameters['script'])
all_paths = []
for path_i in input_parameters['tumor_bam'], input_parameters[
'normal_bam'], input_parameters[
'genome_reference'], input_parameters[
'output_directory'], input_parameters[
'inclusion_region'], input_parameters['dbsnp_gz']:
if path_i:
all_paths.append(path_i)
container_line, fileDict = container.container_params(
input_parameters['lofreq_image'],
tech=tech,
files=all_paths,
extra_args=input_parameters['extra_docker_options'])
# Mounted paths for all the input files and output directory:
mounted_genome_reference = fileDict[
input_parameters['genome_reference']]['mount_path']
mounted_tumor_bam = fileDict[input_parameters['tumor_bam']]['mount_path']
mounted_normal_bam = fileDict[input_parameters['normal_bam']]['mount_path']
mounted_outdir = fileDict[
input_parameters['output_directory']]['mount_path']
mounted_inclusion = fileDict[
input_parameters['inclusion_region']]['mount_path']
mounted_dbsnp_gz = fileDict[input_parameters['dbsnp_gz']]['mount_path']
with open(outfile, 'w') as out:
out.write("#!/bin/bash\n\n")
out.write(f'#$ -o {logdir}\n')
out.write(f'#$ -e {logdir}\n')
out.write('#$ -S /bin/bash\n')
out.write('#$ -l h_vmem={}\n'.format(input_parameters['MEM']))
out.write('set -e\n\n')
out.write('echo -e "Start at `date +"%Y/%m/%d %H:%M:%S"`" 1>&2\n\n')
out.write(f'{container_line} \\\n')
out.write('lofreq somatic \\\n')
out.write('-t {} \\\n'.format(mounted_tumor_bam))
out.write('-n {} \\\n'.format(mounted_normal_bam))
out.write('--call-indels \\\n')
out.write('-l {} \\\n'.format(mounted_inclusion))
out.write('-f {} \\\n'.format(mounted_genome_reference))
out.write('-o {}/{} \\\n'.format(mounted_outdir,
input_parameters['out_prefix']))
if input_parameters['lofreq_arguments']:
out.write('{} \\\n'.format(input_parameters['lofreq_arguments']))
out.write('-d {}\n'.format(mounted_dbsnp_gz))
out.write('\necho -e "Done at `date +"%Y/%m/%d %H:%M:%S"`" 1>&2\n')
# "Run" the script that was generated
command_line = '{} {}'.format(input_parameters['action'], outfile)
returnCode = subprocess.call(command_line, shell=True)
return outfile
def tumor_normal(input_parameters, tech='docker'):
for param_i in DEFAULT_PARAMS:
if param_i not in input_parameters:
input_parameters[param_i] = DEFAULT_PARAMS[param_i]
# The following are required:
assert os.path.exists(input_parameters['normal_bam'])
assert os.path.exists(input_parameters['tumor_bam'])
assert os.path.exists(input_parameters['genome_reference'])
assert os.path.exists(input_parameters['reference_dict'])
logdir = os.path.join(input_parameters['output_directory'], 'logs')
outfile = os.path.join(logdir, input_parameters['script'])
all_paths = []
for path_i in input_parameters['normal_bam'], input_parameters[
'tumor_bam'], input_parameters[
'genome_reference'], input_parameters[
'output_directory'], input_parameters[
'inclusion_region'], input_parameters[
'reference_dict']:
if path_i:
all_paths.append(path_i)
container_line, fileDict = container.container_params(
input_parameters['scalpel_image'],
tech=tech,
files=all_paths,
extra_args=input_parameters['extra_docker_options'])
# Mounted paths for all the input files and output directory:
mounted_genome_reference = fileDict[
input_parameters['genome_reference']]['mount_path']
mounted_tumor_bam = fileDict[input_parameters['tumor_bam']]['mount_path']
mounted_normal_bam = fileDict[input_parameters['normal_bam']]['mount_path']
mounted_outdir = fileDict[
input_parameters['output_directory']]['mount_path']
mounted_reference_dict = fileDict[
input_parameters['reference_dict']]['mount_path']
mounted_inclusion = fileDict[
input_parameters['inclusion_region']]['mount_path']
with open(outfile, 'w') as out:
out.write("#!/bin/bash\n\n")
out.write(f'#$ -o {logdir}\n')
out.write(f'#$ -e {logdir}\n')
out.write('#$ -S /bin/bash\n')
out.write('#$ -l h_vmem={}\n'.format(input_parameters['MEM']))
out.write('set -e\n\n')
out.write('echo -e "Start at `date +"%Y/%m/%d %H:%M:%S"`" 1>&2\n\n')
out.write(f'{container_line} bash -c \\\n')
out.write('"/opt/scalpel/scalpel-discovery --somatic \\\n')
out.write('--ref {} \\\n'.format(mounted_genome_reference))
out.write('--bed {} \\\n'.format(mounted_inclusion))
out.write('--normal {} \\\n'.format(mounted_normal_bam))
out.write('--tumor {} \\\n'.format(mounted_tumor_bam))
out.write('--window 600 \\\n')
if input_parameters['scalpel_two_pass']:
out.write('--two-pass \\\n')
if input_parameters['scalpel_discovery_arguments']:
out.write('{} \\\n'.format(
DISCOVERY_ARGS=input_parameters['scalpel_discovery_arguments'])
)
out.write('--dir {}/scalpel && \\\n'.format(mounted_outdir))
out.write('/opt/scalpel/scalpel-export --somatic \\\n')
out.write(
'--db {}/scalpel/main/somatic.db.dir \\\n'.format(mounted_outdir))
out.write('--ref {} \\\n'.format(mounted_genome_reference))
out.write('--bed {} \\\n'.format(mounted_inclusion))
out.write('{} \\\n'.format(
input_parameters['scalpel_export_arguments']))
out.write('> {}/scalpel/scalpel.vcf"\n\n'.format(mounted_outdir))
out.write(f'{container_line} bash -c \\\n')
out.write(
'"cat {}/scalpel/scalpel.vcf | /opt/vcfsorter.pl {} - \\\n'.format(
mounted_outdir, mounted_reference_dict))
out.write('> {}/{}\"\n'.format(mounted_outdir,
input_parameters['outfile']))
out.write('\necho -e "Done at `date +"%Y/%m/%d %H:%M:%S"`" 1>&2\n')
# "Run" the script that was generated
command_line = '{} {}'.format(input_parameters['action'], outfile)
returnCode = subprocess.call(command_line, shell=True)
return outfile
def picard(inbams,
outbam,
tech='docker',
input_parameters={},
remove_inbams=False):
for param_i in DEFAULT_PARAMS:
if param_i not in input_parameters:
input_parameters[param_i] = DEFAULT_PARAMS[param_i]
logdir = os.path.join(input_parameters['output_directory'], 'logs')
outfile = os.path.join(logdir, input_parameters['script'])
all_paths = list(inbams) + [
outbam,
]
merge_line, fileDict = container.container_params(
input_parameters['picard_image'],
tech=tech,
files=all_paths,
extra_args=input_parameters['extra_docker_options'])
mounted_outbam = fileDict[outbam]['mount_path']
infile_string = ''
for file_i in inbams:
infile_string = infile_string + 'I={} '.format(
fileDict[file_i]['mount_path'])
picard_index_file = re.sub(r'm$', 'i', outbam)
if outbam.endswith('.bam'):
samtools_index_file = outbam + '.bai'
elif outbam.endswith('.cram'):
samtools_index_file = outbam + '.crai'
else:
raise Exception('Output file {} seems wrong.'.format(outbam))
with open(outfile, 'w') as out:
out.write("#!/bin/bash\n\n")
out.write(f'#$ -o {logdir}\n')
out.write(f'#$ -e {logdir}\n')
out.write('#$ -S /bin/bash\n')
out.write('#$ -l h_vmem={}G\n'.format(input_parameters['MEM']))
out.write('set -e\n\n')
out.write(
'echo -e "Start at `date +"%Y/%m/%d %H:%M:%S"`" 1>&2\n\n'
) # Do not change this: picard_fractional uses this to end the copying.
out.write(f'{merge_line} \\\n')
out.write(
'java -Xmx{}G -jar /opt/picard.jar MergeSamFiles {} {} ASSUME_SORTED=true CREATE_INDEX=true O={}\n\n'
.format(input_parameters['MEM'], infile_string,
input_parameters['extra_picard_arguments'],
mounted_outbam))
if remove_inbams:
out.write('rm {}\n\n'.format(' '.join(inbams)))
out.write('mv {} {}\n\n'.format(picard_index_file,
samtools_index_file))
out.write('\necho -e "Done at `date +"%Y/%m/%d %H:%M:%S"`" 1>&2\n')
# "Run" the script that was generated
command_line = '{} {}'.format(input_parameters['action'], outfile)
returnCode = subprocess.call(command_line, shell=True)
return outfile
def picard(input_parameters, tech='docker'):
for param_i in DEFAULT_PARAMS:
if param_i not in input_parameters:
input_parameters[param_i] = DEFAULT_PARAMS[param_i]
#
logdir = os.path.join(input_parameters['output_directory'], 'logs')
outfile = os.path.join(logdir, input_parameters['script'])
all_paths = []
for path_i in input_parameters['output_directory'], input_parameters[
'in_bam']:
if path_i:
all_paths.append(path_i)
markdup_line, fileDict = container.container_params(
input_parameters['picard_image'],
tech=tech,
files=all_paths,
extra_args=input_parameters['extra_docker_options'])
samtools_line, stDict = container.container_params(
input_parameters['samtools_image'], tech, [
input_parameters['output_directory'],
], input_parameters['extra_docker_options'])
# Mounted paths for all the input files and output directory:
mounted_outdir = fileDict[
input_parameters['output_directory']]['mount_path']
mounted_inbam = fileDict[input_parameters['in_bam']]['mount_path']
tempdir = uuid.uuid4().hex
with open(outfile, 'w') as out:
out.write("#!/bin/bash\n\n")
out.write(f'#$ -o {logdir}\n')
out.write(f'#$ -e {logdir}\n')
out.write('#$ -S /bin/bash\n')
out.write('#$ -l h_vmem={}G\n'.format(input_parameters['MEM']))
out.write('set -e\n\n')
out.write(
'echo -e "Start at `date +"%Y/%m/%d %H:%M:%S"`" 1>&2\n\n'
) # Do not change this: fractional uses this to end the copying.
out.write('mkdir -p {}/{}\n\n'.format(
input_parameters['output_directory'], tempdir))
out.write(f'{markdup_line} \\\n')
out.write(
'java -Xmx{}G -jar /opt/picard.jar MarkDuplicatesWithMateCigar \\\n'
.format(input_parameters['MEM']))
out.write('I={} \\\n'.format(mounted_inbam))
out.write('M={}/{} \\\n'.format(
mounted_outdir,
re.sub(
r'\.(bam|cram)', '',
fileDict[input_parameters['in_bam']]['filename'] +
'.markdup')))
out.write('ASSUME_SORT_ORDER=coordinate \\\n')
out.write('TMP_DIR={}/{} \\\n'.format(mounted_outdir, tempdir))
out.write('MINIMUM_DISTANCE=1000 \\\n')
out.write('O={}/{}\n\n'.format(mounted_outdir,
input_parameters['out_bam']))
if input_parameters['index_bam']:
out.write(f'{samtools_line} \\\n')
out.write('samtools index -@{} {}/{}\n\n'.format(
input_parameters['threads'],
stDict[input_parameters['output_directory']]['mount_path'],
input_parameters['out_bam']))
out.write('rm -r {}/{}\n'.format(input_parameters['output_directory'],
tempdir))
out.write(
'\necho -e "Done at `date +"%Y/%m/%d %H:%M:%S"`" 1>&2\n'
) # Do not change this: fractional uses this to end the copying.
# "Run" the script that was generated
command_line = '{} {}'.format(input_parameters['action'], outfile)
returnCode = subprocess.call(command_line, shell=True)
return outfile
def fractional(bed, input_parameters, tech='docker'):
for param_i in DEFAULT_PARAMS:
if param_i not in input_parameters:
input_parameters[param_i] = DEFAULT_PARAMS[param_i]
outdir = str(Path(bed).absolute().parent)
logdir = os.path.join(outdir, 'logs')
outfile = os.path.join(logdir, 'markdup_fractional.{}.cmd'.format(ts))
os.makedirs(logdir, exist_ok=True)
sambam_line, stDict = container.container_params(
input_parameters['sambamba_image'], tech, [
input_parameters['in_bam'],
bed,
], input_parameters['extra_docker_options'])
mounted_inbam = stDict[input_parameters['in_bam']]['mount_path']
mounted_bed = stDict[bed]['mount_path']
mounted_outdir = stDict[bed]['mount_dir']
temp_split_bam = uuid.uuid4().hex + '.bam'
split_deduped_bam = uuid.uuid4().hex + '.bam'
with open(outfile, 'w') as out:
out.write("#!/bin/bash\n\n")
out.write(f'#$ -o {logdir}\n')
out.write(f'#$ -e {logdir}\n')
out.write('#$ -S /bin/bash\n')
out.write('#$ -l h_vmem={}G\n'.format(input_parameters['MEM']))
out.write('set -e\n\n')
out.write('echo -e "Start at `date +"%Y/%m/%d %H:%M:%S"`" 1>&2\n\n')
out.write(f'{sambam_line} \\\n')
out.write('sambamba view -L {} -t {} -f bam -o {} {}\n\n'.format(
mounted_bed, 1, os.path.join(mounted_outdir, temp_split_bam),
mounted_inbam))
fractional_parameters = copy(input_parameters)
fractional_parameters['output_directory'] = outdir
fractional_parameters['in_bam'] = os.path.join(outdir, temp_split_bam)
fractional_parameters['out_bam'] = split_deduped_bam
fractional_parameters['script'] = 'to_be_deleted.{}.cmd'.format(ts)
fractional_parameters['index_bam'] = False
if input_parameters['software'] == 'picard':
dedup_script = picard(fractional_parameters, tech)
elif input_parameters['software'] == 'sambamba':
fractional_parameters['threads'] = 2
dedup_script = sambamba(fractional_parameters, tech)
with open(os.path.join(logdir,
fractional_parameters['script'])) as dedup:
line_i = dedup.readline()
while not line_i.startswith('echo -e "Start'):
line_i = dedup.readline()
while not line_i.startswith('echo -e "Done'):
out.write(line_i)
line_i = dedup.readline()
out.write('rm {}\n'.format(os.path.join(outdir, temp_split_bam)))
out.write('\necho -e "Done at `date +"%Y/%m/%d %H:%M:%S"`" 1>&2\n')
# "Run" the script that was generated
command_line = '{} {}'.format(input_parameters['action'], outfile)
returnCode = subprocess.call(command_line, shell=True)
return outfile, os.path.join(outdir, split_deduped_bam)
def sambamba(input_parameters, tech='docker'):
for param_i in DEFAULT_PARAMS:
if param_i not in input_parameters:
input_parameters[param_i] = DEFAULT_PARAMS[param_i]
#
logdir = os.path.join(input_parameters['output_directory'], 'logs')
outfile = os.path.join(logdir, input_parameters['script'])
all_paths = []
for path_i in input_parameters['output_directory'], input_parameters[
'in_bam']:
if path_i:
all_paths.append(path_i)
markdup_line, fileDict = container.container_params(
input_parameters['sambamba_image'],
tech=tech,
files=all_paths,
extra_args=input_parameters['extra_docker_options'])
# Mounted paths for all the input files and output directory:
mounted_outdir = fileDict[
input_parameters['output_directory']]['mount_path']
mounted_inbam = fileDict[input_parameters['in_bam']]['mount_path']
tempdir = uuid.uuid4().hex
with open(outfile, 'w') as out:
out.write("#!/bin/bash\n\n")
out.write(f'#$ -o {logdir}\n')
out.write(f'#$ -e {logdir}\n')
out.write('#$ -S /bin/bash\n')
out.write('#$ -l h_vmem={}G\n'.format(input_parameters['MEM']))
out.write('set -e\n\n')
out.write(
'echo -e "Start at `date +"%Y/%m/%d %H:%M:%S"`" 1>&2\n\n'
) # Do not change this: fractional uses this to end the copying.
out.write('mkdir -p {}/{}\n\n'.format(
input_parameters['output_directory'], tempdir))
out.write(f'{markdup_line} \\\n')
out.write('sambamba markdup -t {} --tmpdir {} {} {}\n\n'.format(
input_parameters['threads'], os.path.join(mounted_outdir, tempdir),
mounted_inbam,
os.path.join(mounted_outdir, input_parameters['out_bam'])))
out.write('rm -r {}/{}\n'.format(input_parameters['output_directory'],
tempdir))
out.write(
'\necho -e "Done at `date +"%Y/%m/%d %H:%M:%S"`" 1>&2\n'
) # Do not change this: fractional uses this to end the copying.
# "Run" the script that was generated
command_line = '{} {}'.format(input_parameters['action'], outfile)
returnCode = subprocess.call(command_line, shell=True)
return outfile
def tumor_normal(input_parameters=DEFAULT_PARAMS, tech='docker'):
for param_i in DEFAULT_PARAMS:
if param_i not in input_parameters:
input_parameters[param_i] = DEFAULT_PARAMS[param_i]
# The following are required:
assert os.path.exists(input_parameters['normal_bam'])
assert os.path.exists(input_parameters['tumor_bam'])
assert os.path.exists(input_parameters['genome_reference'])
assert os.path.exists(input_parameters['dbsnp_gz'])
assert os.path.exists(input_parameters['dbsnp_gz'] + '.tbi')
logdir = os.path.join(input_parameters['output_directory'], 'logs')
outfile = os.path.join(logdir, input_parameters['script'])
all_paths = []
for path_i in input_parameters['normal_bam'], input_parameters[
'tumor_bam'], input_parameters[
'genome_reference'], input_parameters[
'output_directory'], input_parameters[
'inclusion_region'], input_parameters['dbsnp_gz']:
if path_i:
all_paths.append(path_i)
container_line, fileDict = container.container_params(
input_parameters['muse_image'],
tech=tech,
files=all_paths,
extra_args=input_parameters['extra_docker_options'])
# Mounted paths for all the input files and output directory:
mounted_genome_reference = fileDict[
input_parameters['genome_reference']]['mount_path']
mounted_tumor_bam = fileDict[input_parameters['tumor_bam']]['mount_path']
mounted_normal_bam = fileDict[input_parameters['normal_bam']]['mount_path']
mounted_outdir = fileDict[
input_parameters['output_directory']]['mount_path']
mounted_dbsnp_gz = fileDict[input_parameters['dbsnp_gz']]['mount_path']
with open(outfile, 'w') as out:
out.write("#!/bin/bash\n\n")
out.write(f'#$ -o {logdir}\n')
out.write(f'#$ -e {logdir}\n')
out.write('#$ -S /bin/bash\n')
out.write('#$ -l h_vmem={}\n'.format(input_parameters['MEM']))
out.write('set -e\n\n')
out.write('echo -e "Start at `date +"%Y/%m/%d %H:%M:%S"`" 1>&2\n\n')
out.write(
'cat {} | awk -F "\\t" \'{{print $1 "\\t" $2 "\\t" $3}}\' > {}/bed_3columns.bed\n\n'
.format(input_parameters['inclusion_region'],
input_parameters['output_directory']))
out.write(f'{container_line} \\\n')
out.write('MuSEv1.0rc_submission_c039ffa call \\\n')
out.write('-O {}/MuSE \\\n'.format(mounted_outdir))
out.write('-l {}/bed_3columns.bed \\\n'.format(mounted_outdir))
out.write('-f {} \\\n'.format(mounted_genome_reference))
out.write('{} \\\n'.format(mounted_tumor_bam))
out.write('{}\n\n'.format(mounted_normal_bam))
out.write(f'{container_line} \\\n')
out.write('MuSEv1.0rc_submission_c039ffa sump \\\n')
out.write('-I {}/MuSE.MuSE.txt \\\n'.format(mounted_outdir))
if input_parameters['exome']:
out.write('-E \\\n')
else:
out.write('-G \\\n')
if input_parameters['muse_arguments']:
out.write('{} \\\n'.format(
EXTRA_ARGS=input_parameters['muse_arguments']))
out.write('-O {}/{} \\\n'.format(mounted_outdir,
input_parameters['outfile']))
out.write('-D {}\n'.format(mounted_dbsnp_gz))
out.write('\necho -e "Done at `date +"%Y/%m/%d %H:%M:%S"`" 1>&2\n')
# "Run" the script that was generated
command_line = '{} {}'.format(input_parameters['action'], outfile)
returnCode = subprocess.call(command_line, shell=True)
return outfile
def tumor_normal(input_parameters=DEFAULT_PARAMS, tech='docker'):
for param_i in DEFAULT_PARAMS:
if param_i not in input_parameters:
input_parameters[param_i] = DEFAULT_PARAMS[param_i]
# The following are required:
assert os.path.exists(input_parameters['normal_bam'])
assert os.path.exists(input_parameters['tumor_bam'])
assert os.path.exists(input_parameters['genome_reference'])
logdir = os.path.join(input_parameters['output_directory'], 'logs')
outfile = os.path.join(logdir, input_parameters['script'])
all_paths = []
for path_i in input_parameters['normal_bam'], input_parameters[
'tumor_bam'], input_parameters[
'genome_reference'], input_parameters[
'output_directory'], input_parameters['inclusion_region']:
if path_i:
all_paths.append(path_i)
container_line, fileDict = container.container_params(
input_parameters['varscan2_image'],
tech=tech,
files=all_paths,
extra_args=input_parameters['extra_docker_options'])
mpileine_line, plDict = container.container_params(
'lethalfang/samtools:1.7',
tech=tech,
files=all_paths,
extra_args=input_parameters['extra_docker_options'])
# Mounted paths for all the input files and output directory:
mounted_genome_reference = fileDict[
input_parameters['genome_reference']]['mount_path']
mounted_tumor_bam = fileDict[input_parameters['tumor_bam']]['mount_path']
mounted_normal_bam = fileDict[input_parameters['normal_bam']]['mount_path']
mounted_outdir = fileDict[
input_parameters['output_directory']]['mount_path']
# Mounted paths for mpileup dockers
pl_genome_reference = plDict[
input_parameters['genome_reference']]['mount_path']
pl_tumor_bam = plDict[input_parameters['tumor_bam']]['mount_path']
pl_normal_bam = plDict[input_parameters['normal_bam']]['mount_path']
pl_outdir = plDict[input_parameters['output_directory']]['mount_path']
if input_parameters['inclusion_region']:
selector_text = '-l {}'.format(
plDict[input_parameters['inclusion_region']]['mount_path'])
else:
selector_text = ''
if input_parameters['minimum_VAF']:
minVAF = input_parameters['minimum_VAF']
outname = re.sub(r'\.[a-zA-Z]+$', '', input_parameters['outfile'])
with open(outfile, 'w') as out:
out.write("#!/bin/bash\n\n")
out.write(f'#$ -o {logdir}\n')
out.write(f'#$ -e {logdir}\n')
out.write('#$ -S /bin/bash\n')
out.write('#$ -l h_vmem={}\n'.format(input_parameters['MEM']))
out.write('set -e\n\n')
out.write('echo -e "Start at `date +"%Y/%m/%d %H:%M:%S"`" 1>&2\n\n')
out.write(f'{mpileine_line} bash -c \\\n')
out.write('"samtools mpileup \\\n')
out.write(
'-B -q {minMQ} -Q {minBQ} {extra_pileup_arguments} {selector_text} -f \\\n'
.format(minMQ=input_parameters['min_MQ'],
minBQ=input_parameters['min_BQ'],
extra_pileup_arguments=input_parameters[
'varscan_pileup_arguments'],
selector_text=selector_text))
out.write('{} \\\n'.format(pl_genome_reference))
out.write('{} \\\n'.format(pl_normal_bam))
out.write('> {}/normal.pileup"\n\n'.format(pl_outdir))
out.write(f'{mpileine_line} bash -c \\\n')
out.write('"samtools mpileup \\\n')
out.write(
'-B -q {minMQ} -Q {minBQ} {extra_pileup_arguments} {selector_text} -f \\\n'
.format(minMQ=input_parameters['min_MQ'],
minBQ=input_parameters['min_BQ'],
extra_pileup_arguments=input_parameters[
'varscan_pileup_arguments'],
selector_text=selector_text))
out.write('{} \\\n'.format(pl_genome_reference))
out.write('{} \\\n'.format(pl_tumor_bam))
out.write('> {}/tumor.pileup"\n\n'.format(pl_outdir))
out.write(f'{container_line} \\\n')
out.write('java -Xmx{} -jar /VarScan2.3.7.jar somatic \\\n'.format(
input_parameters['MEM']))
out.write('{}/normal.pileup \\\n'.format(mounted_outdir))
out.write('{}/tumor.pileup \\\n'.format(mounted_outdir))
out.write('{}/{} {} --output-vcf 1 --min-var-freq {}\n\n'.format(
mounted_outdir, outname, input_parameters['varscan_arguments'],
input_parameters['minimum_VAF']))
out.write(f'{container_line} \\\n')
out.write(
'java -Xmx{} -jar /VarScan2.3.7.jar processSomatic \\\n'.format(
input_parameters['MEM']))
out.write('{}/{}.snp.vcf\n\n'.format(mounted_outdir, outname))
out.write(f'{container_line} \\\n')
out.write(
'java -Xmx{} -jar /VarScan2.3.7.jar somaticFilter \\\n'.format(
input_parameters['MEM']))
out.write('{}/{}.snp.Somatic.hc.vcf \\\n'.format(
mounted_outdir, outname))
out.write('-indel-file {}/{}.indel.vcf \\\n'.format(
mounted_outdir, outname))
out.write('-output-file {}/{}.snp.Somatic.hc.filter.vcf\n\n'.format(
mounted_outdir, outname))
out.write('rm {}/normal.pileup\n'.format(
input_parameters['output_directory']))
out.write('rm {}/tumor.pileup\n'.format(
input_parameters['output_directory']))
out.write('\necho -e "Done at `date +"%Y/%m/%d %H:%M:%S"`" 1>&2\n')
# "Run" the script that was generated
command_line = '{} {}'.format(input_parameters['action'], outfile)
returnCode = subprocess.call(command_line, shell=True)
return outfile
def tumor_normal(input_parameters=DEFAULT_PARAMS, tech='docker'):
for param_i in DEFAULT_PARAMS:
if param_i not in input_parameters:
input_parameters[param_i] = DEFAULT_PARAMS[param_i]
# The following are required:
assert os.path.exists(input_parameters['normal_bam'])
assert os.path.exists(input_parameters['tumor_bam'])
assert os.path.exists(input_parameters['genome_reference'])
assert os.path.exists(input_parameters['reference_dict'])
logdir = os.path.join(input_parameters['output_directory'], 'logs')
outfile = os.path.join(logdir, input_parameters['script'])
all_paths = []
for path_i in input_parameters['normal_bam'], input_parameters[
'tumor_bam'], input_parameters[
'genome_reference'], input_parameters[
'output_directory'], input_parameters['reference_dict']:
if path_i:
all_paths.append(path_i)
container_line, fileDict = container.container_params(
input_parameters['jsm2_image'],
tech=tech,
files=all_paths,
extra_args=input_parameters['extra_docker_options'])
# Mounted paths for all the input files and output directory:
mounted_genome_reference = fileDict[
input_parameters['genome_reference']]['mount_path']
mounted_tumor_bam = fileDict[input_parameters['tumor_bam']]['mount_path']
mounted_normal_bam = fileDict[input_parameters['normal_bam']]['mount_path']
mounted_outdir = fileDict[
input_parameters['output_directory']]['mount_path']
mounted_reference_dict = fileDict[
input_parameters['reference_dict']]['mount_path']
with open(outfile, 'w') as out:
out.write("#!/bin/bash\n\n")
out.write(f'#$ -o {logdir}\n')
out.write(f'#$ -e {logdir}\n')
out.write('#$ -S /bin/bash\n')
out.write('#$ -l h_vmem={}\n'.format(input_parameters['MEM']))
out.write('set -e\n\n')
out.write('echo -e "Start at `date +"%Y/%m/%d %H:%M:%S"`" 1>&2\n')
out.write('\n')
out.write(f'{container_line} \\\n')
out.write(
'/opt/JointSNVMix-0.7.5/build/scripts-2.7/jsm.py train joint_snv_mix_two \\\n'
)
out.write('--convergence_threshold {} \\\n'.format(
input_parameters['converge_threshold']))
out.write('--skip_size {} \\\n'.format(input_parameters['skip_size']))
if input_parameters['jsm_train_arguments']:
out.write('{} \\\n'.format(
input_parameters['jsm_train_arguments']))
out.write('{} \\\n'.format(mounted_genome_reference))
out.write('{} \\\n'.format(mounted_normal_bam))
out.write('{} \\\n'.format(mounted_tumor_bam))
out.write('/opt/JointSNVMix-0.7.5/config/joint_priors.cfg \\\n')
out.write('/opt/JointSNVMix-0.7.5/config/joint_params.cfg \\\n')
out.write('{}/jsm.parameter.cfg\n'.format(mounted_outdir))
out.write('\n')
out.write('echo -e \'##fileformat=VCFv4.1\' > {}/{}\n'.format(
input_parameters['output_directory'], input_parameters['outfile']))
out.write(
'echo -e \'##INFO=<ID=AAAB,Number=1,Type=Float,Description="Probability of Joint Genotype AA in Normal and AB in Tumor">\' >> {}/{}\n'
.format(input_parameters['output_directory'],
input_parameters['outfile']))
out.write(
'echo -e \'##INFO=<ID=AABB,Number=1,Type=Float,Description="Probability of Joint Genotype AA in Normal and BB in Tumor">\' >> {}/{}\n'
.format(input_parameters['output_directory'],
input_parameters['outfile']))
out.write(
'echo -e \'##FORMAT=<ID=GT,Number=1,Type=String,Description="Genotype">\' >> {}/{}\n'
.format(input_parameters['output_directory'],
input_parameters['outfile']))
out.write(
'echo -e \'##FORMAT=<ID=RD,Number=1,Type=Integer,Description="Depth of reference-supporting bases (reads1)">\' >> {}/{}\n'
.format(input_parameters['output_directory'],
input_parameters['outfile']))
out.write(
'echo -e \'##FORMAT=<ID=AD,Number=1,Type=Integer,Description="Depth of variant-supporting bases (reads2)">\' >> {}/{}\n'
.format(input_parameters['output_directory'],
input_parameters['outfile']))
out.write(
'echo -e \'#CHROM\\tPOS\\tID\\tREF\\tALT\\tQUAL\\tFILTER\\tINFO\\tFORMAT\\tNORMAL\\tTUMOR\' >> {}/{}\n'
.format(input_parameters['output_directory'],
input_parameters['outfile']))
out.write('\n')
out.write(f'{container_line} bash -c \\\n')
out.write(
'"/opt/JointSNVMix-0.7.5/build/scripts-2.7/jsm.py classify joint_snv_mix_two \\\n'
)
if input_parameters['jsm_classify_arguments']:
out.write('{} \\\n'.format(
input_parameters['jsm_classify_arguments']))
out.write('{} \\\n'.format(mounted_genome_reference))
out.write('{} \\\n'.format(mounted_normal_bam))
out.write('{} \\\n'.format(mounted_tumor_bam))
out.write("{}/jsm.parameter.cfg \\\n".format(mounted_outdir))
out.write(
'/dev/stdout | awk -F \'\\t\' \'NR!=1 && \\$4!=\\"N\\" && \\$10+\\$11>=0.95\' | \\\n'
)
out.write(
'awk -F \'\\t\' \'{print \\$1 \\"\\t\\" \\$2 \\"\\t.\\t\\" \\$3 \\"\\t\\" \\$4 \\"\\t.\\t.\\tAAAB=\\" \\$10 \\";AABB=\\" \\$11 \\"\\tRD:AD\\t\\" \\$5 \\":\\" \\$6 \\"\\t\\" \\$7 \\":\\" \\$8}\' \\\n'
)
out.write('| /opt/vcfsorter.pl {} - >> {}/{}"\n\n'.format(
mounted_reference_dict, mounted_outdir,
input_parameters['outfile']))
if input_parameters['threads'] > 1:
bedtool_line, outdir_i = container.container_params(
'lethalfang/bedtools:2.26.0', tech,
(input_parameters['output_directory'], ))
mounted_bed_outdir = outdir_i[
input_parameters['output_directory']]['mount_path']
out.write('\n\ni=1\n')
out.write('while [[ $i -le {} ]]\n'.format(
input_parameters['threads']))
out.write('do\n')
out.write(
' {DOCKER_LINE} bash -c "bedtools intersect -a {OUTDIR}/{OUTVCF} -b {OUTDIR}/${{i}}/${{i}}.bed -header | uniq > {OUTDIR}/${{i}}/{OUTVCF}"\n'
.format(DOCKER_LINE=bedtool_line,
OUTDIR=mounted_bed_outdir,
OUTVCF=input_parameters['outfile']))
out.write(' i=$(( $i + 1 ))\n')
out.write('done\n')
out.write('\necho -e "Done at `date +"%Y/%m/%d %H:%M:%S"`" 1>&2\n')
# "Run" the script that was generated
command_line = '{} {}'.format(input_parameters['action'], outfile)
returnCode = subprocess.call(command_line, shell=True)
return outfile
def trimmomatic(input_parameters, tech='docker'):
if input_parameters['in_fastq2']:
paired_end = True
else:
paired_end = False
for param_i in DEFAULT_PARAMS:
if param_i not in input_parameters:
input_parameters[param_i] = DEFAULT_PARAMS[param_i]
#
logdir = os.path.join(input_parameters['output_directory'], 'logs')
outfile = os.path.join(logdir, input_parameters['script'])
all_paths = []
for path_i in input_parameters['output_directory'], input_parameters[
'in_fastq1'], input_parameters['in_fastq2']:
if path_i:
all_paths.append(path_i)
trim_line, fileDict = container.container_params(
input_parameters['trimmomaticImage'],
tech=tech,
files=all_paths,
extra_args=input_parameters['extra_docker_options'])
# Mounted paths for all the input files and output directory:
mounted_outdir = fileDict[
input_parameters['output_directory']]['mount_path']
mounted_fq1 = fileDict[input_parameters['in_fastq1']]['mount_path']
mounted_fq2 = fileDict[input_parameters['in_fastq2']]['mount_path']
temporary_files = []
with open(outfile, 'w') as out:
out.write("#!/bin/bash\n\n")
out.write(f'#$ -o {logdir}\n')
out.write(f'#$ -e {logdir}\n')
out.write('#$ -S /bin/bash\n')
out.write('#$ -l h_vmem={}G\n'.format(input_parameters['MEM']))
out.write('set -e\n\n')
out.write('echo -e "Start at `date +"%Y/%m/%d %H:%M:%S"`" 1>&2\n\n')
out.write(f'{trim_line} \\\n')
out.write(
'java -Xmx{}G -jar /opt/Trimmomatic/trimmomatic.jar \\\n'.format(
input_parameters['MEM']))
if paired_end:
out.write('PE -threads {} -phred33 \\\n'.format(
input_parameters['threads']))
out.write(
'{FQ1} {FQ2} {DIR}/{PAIR1} {DIR}/{UNPAIR1} {DIR}/{PAIR2} {DIR}/{UNPAIR2} \\\n'
.format(
FQ1=mounted_fq1,
FQ2=mounted_fq2,
DIR=mounted_outdir,
PAIR1=input_parameters['out_fastq1_name'],
PAIR2=input_parameters['out_fastq2_name'],
UNPAIR1='unpaired.' + input_parameters['out_fastq1_name'],
UNPAIR2='unpaired.' + input_parameters['out_fastq2_name']))
else:
out.write('SE -threads {} -phred33 \\\n'.format(
input_parameters['threads']))
out.write('{FQ1} {DIR}/{PAIR1} \\\n'.format(
FQ1=mounted_fq1,
DIR=mounted_outdir,
PAIR1=input_parameters['out_fastq1_name']))
out.write(
'ILLUMINACLIP:{ADAPTER}:2:30:10 LEADING:3 TRAILING:3 SLIDINGWINDOW:4:15 MINLEN:{MINLEN}\n'
.format(ADAPTER=input_parameters['adapter'],
MINLEN=input_parameters['minimum_length']))
# Remove untrimmed files:
if input_parameters['remove_untrimmed']:
out.write('\n')
out.write('rm {}\n'.format(input_parameters['in_fastq1']))
if input_parameters['in_fastq2']:
out.write('rm {}\n'.format(input_parameters['in_fastq2']))
out.write('\necho -e "Done at `date +"%Y/%m/%d %H:%M:%S"`" 1>&2\n')
# "Run" the script that was generated
command_line = '{} {}'.format(input_parameters['action'], outfile)
returnCode = subprocess.call(command_line, shell=True)
return outfile
def tumor_normal(input_parameters=DEFAULT_PARAMS, tech='docker'):
for param_i in DEFAULT_PARAMS:
if param_i not in input_parameters:
input_parameters[param_i] = DEFAULT_PARAMS[param_i]
# The following are required:
assert os.path.exists(input_parameters['normal_bam'])
assert os.path.exists(input_parameters['tumor_bam'])
assert os.path.exists(input_parameters['genome_reference'])
logdir = os.path.join(input_parameters['output_directory'], 'logs')
outfile = os.path.join(logdir, input_parameters['script'])
all_paths = []
for path_i in input_parameters['normal_bam'], input_parameters[
'tumor_bam'], input_parameters[
'genome_reference'], input_parameters[
'output_directory'], input_parameters['inclusion_region']:
if path_i:
all_paths.append(path_i)
container_line, fileDict = container.container_params(
input_parameters['mutect2_image'],
tech=tech,
files=all_paths,
extra_args=input_parameters['extra_docker_options'])
tumor_name_line, tumor_bam = container.container_params(
'lethalfang/samtools:1.7', tech, (input_parameters['tumor_bam'], ))
normal_name_line, normal_bam = container.container_params(
'lethalfang/samtools:1.7', tech, (input_parameters['normal_bam'], ))
# Resolve mounted paths
mounted_genome_reference = fileDict[
input_parameters['genome_reference']]['mount_path']
mounted_tumor_bam = fileDict[input_parameters['tumor_bam']]['mount_path']
mounted_normal_bam = fileDict[input_parameters['normal_bam']]['mount_path']
mounted_outdir = fileDict[
input_parameters['output_directory']]['mount_path']
if input_parameters['inclusion_region']:
mounted_inclusion = fileDict[
input_parameters['inclusion_region']]['mount_path']
with open(outfile, 'w') as out:
out.write("#!/bin/bash\n\n")
out.write(f'#$ -o {logdir}\n')
out.write(f'#$ -e {logdir}\n')
out.write('#$ -S /bin/bash\n')
out.write('#$ -l h_vmem={}\n'.format(input_parameters['MEM']))
out.write('set -e\n\n')
out.write('echo -e "Start at `date +"%Y/%m/%d %H:%M:%S"`" 1>&2\n\n')
tumor_bam_path = tumor_bam[input_parameters['tumor_bam']]['mount_path']
tumor_sample_name_extraction = f'tumor_name=`{tumor_name_line} samtools view -H {tumor_bam_path} | egrep -w \'^@RG\' | grep -Po \'SM:[^\\t$]+\' | sed \'s/SM://\' | uniq | sed -e \'s/[[:space:]]*$//\'`\n'
out.write(tumor_sample_name_extraction)
normal_bam_path = normal_bam[
input_parameters['normal_bam']]['mount_path']
normal_sample_name_extraction = f'normal_name=`{normal_name_line} samtools view -H {normal_bam_path} | egrep -w \'^@RG\' | grep -Po \'SM:[^\\t$]+\' | sed \'s/SM://\' | uniq | sed -e \'s/[[:space:]]*$//\'`\n'
out.write(normal_sample_name_extraction)
out.write('\n')
out.write(f'{container_line} \\\n')
out.write('java -Xmx{} -jar /gatk/gatk.jar Mutect2 \\\n'.format(
input_parameters['MEM']))
out.write(f'--reference {mounted_genome_reference} \\\n')
if input_parameters['inclusion_region']:
out.write('--intervals {} \\\n'.format(mounted_inclusion))
out.write('--input {} \\\n'.format(mounted_tumor_bam))
out.write('--input {} \\\n'.format(mounted_normal_bam))
out.write('--normal-sample ${normal_name} \\\n')
out.write('--tumor-sample ${tumor_name} \\\n')
out.write('--native-pair-hmm-threads {} \\\n'.format(1))
if input_parameters['mutect2_arguments']:
out.write('{} \\\n'.format(input_parameters['mutect2_arguments']))
out.write('--output {}/unfiltered.{}\n\n'.format(
mounted_outdir, input_parameters['outfile']))
out.write(f'{container_line} \\\n')
out.write(
'java -Xmx{} -jar /gatk/gatk.jar FilterMutectCalls \\\n'.format(
input_parameters['MEM']))
out.write('--variant {}/unfiltered.{} \\\n'.format(
mounted_outdir, input_parameters['outfile']))
if input_parameters['mutect2_filter_arguments']:
out.write('{} \\\n'.format(
input_parameters['mutect2_filter_arguments']))
out.write('--output {}/{}\n'.format(mounted_outdir,
input_parameters['outfile']))
out.write('\necho -e "Done at `date +"%Y/%m/%d %H:%M:%S"`" 1>&2\n')
# "Run" the script that was generated
command_line = '{} {}'.format(input_parameters['action'], outfile)
returnCode = subprocess.call(command_line, shell=True)
return outfile
def tumor_normal(input_parameters=DEFAULT_PARAMS, tech='docker' ):
for param_i in DEFAULT_PARAMS:
if param_i not in input_parameters:
input_parameters[param_i] = DEFAULT_PARAMS[param_i]
# The following are required:
assert os.path.exists( input_parameters['normal_bam'] )
assert os.path.exists( input_parameters['tumor_bam'] )
assert os.path.exists( input_parameters['genome_reference'] )
logdir = os.path.join( input_parameters['output_directory'], 'logs' )
outfile = os.path.join( logdir, input_parameters['script'] )
all_paths = []
for path_i in input_parameters['normal_bam'], input_parameters['tumor_bam'], input_parameters['genome_reference'], input_parameters['output_directory'], input_parameters['inclusion_region']:
if path_i:
all_paths.append( path_i )
container_line, fileDict = container.container_params( input_parameters['somaticsniper_image'], tech=tech, files=all_paths, extra_args=input_parameters['extra_docker_options'] )
# Mounted paths for all the input files and output directory:
mounted_genome_reference = fileDict[ input_parameters['genome_reference'] ]['mount_path']
mounted_tumor_bam = fileDict[ input_parameters['tumor_bam'] ]['mount_path']
mounted_normal_bam = fileDict[ input_parameters['normal_bam'] ]['mount_path']
mounted_outdir = fileDict[ input_parameters['output_directory'] ]['mount_path']
if input_parameters['inclusion_region']:
mounted_inclusion = fileDict[ input_parameters['inclusion_region'] ]['mount_path']
with open(outfile, 'w') as out:
out.write( "#!/bin/bash\n\n" )
out.write(f'#$ -o {logdir}\n' )
out.write(f'#$ -e {logdir}\n' )
out.write( '#$ -S /bin/bash\n' )
out.write( '#$ -l h_vmem={}\n'.format( input_parameters['MEM'] ) )
out.write( 'set -e\n\n' )
out.write( 'echo -e "Start at `date +"%Y/%m/%d %H:%M:%S"`" 1>&2\n\n' )
out.write(f'{container_line} \\\n' )
out.write( '/opt/somatic-sniper/build/bin/bam-somaticsniper \\\n' )
out.write( '-q {} -Q {} -s {} -F vcf {} \\\n'.format(input_parameters['min_MQ'], input_parameters['somatic_score'], input_parameters['prior'], input_parameters['somaticsniper_arguments']) )
out.write( '-f {} \\\n'.format(mounted_genome_reference) )
out.write( '{} \\\n'.format(mounted_tumor_bam) )
out.write( '{} \\\n'.format(mounted_normal_bam) )
out.write( '{}/{}\n'.format(mounted_outdir, input_parameters['outfile']) )
if input_parameters['threads'] > 1:
bedtool_line, outdir_i = container.container_params( 'lethalfang/bedtools:2.26.0', tech, (input_parameters['output_directory'], ) )
mounted_bed_outdir = outdir_i[ input_parameters['output_directory'] ]['mount_path']
out.write( '\n\ni=1\n' )
out.write( 'while [[ $i -le {} ]]\n'.format(input_parameters['threads']) )
out.write( 'do\n' )
out.write( ' {DOCKER_LINE} bash -c "bedtools intersect -a {OUTDIR}/{OUTVCF} -b {OUTDIR}/${{i}}/${{i}}.bed -header | uniq > {OUTDIR}/${{i}}/{OUTVCF}"\n'.format(DOCKER_LINE=bedtool_line, OUTDIR=mounted_bed_outdir, OUTVCF=input_parameters['outfile']) )
out.write( ' i=$(( $i + 1 ))\n' )
out.write( 'done\n' )
out.write( '\necho -e "Done at `date +"%Y/%m/%d %H:%M:%S"`" 1>&2\n' )
# "Run" the script that was generated
command_line = '{} {}'.format( input_parameters['action'], outfile )
returnCode = subprocess.call( command_line, shell=True )
return outfile
