def start_ab(args, logger):
'''Perform alignment and bam processing'''
import os
import subprocess
final_bam = args.outbam
# initialize library file from given arguments
library = genobox_modules.initialize_library(args.libfile, args.se,
args.pe1, args.pe2,
args.sample, args.mapq,
args.libs, args.pl)
# start run
if args.sample:
print "--------------------------------------"
print "Processing sample: %s" % args.sample
print "--------------------------------------"
print "Starting alignment"
(bamfiles, library) = start_alignment(args, logger)
print "Starting bam processing"
final_bam = start_bamprocess(library,
genobox_modules.unique(bamfiles.values()),
args.mapq, args.libs, args.tmpdir, args.queue,
final_bam, args.realignment, args.known,
args.fa, args.sample, args.partition, logger)
# remove queuing system outfiles
genobox_modules.rm_files(['run_genobox_*', 'semaphores.*'])
print "Done"
print "--------------------------------------"
def start_ab(args, logger):
'''Perform alignment and bam processing'''
import os
import subprocess
final_bam = args.outbam
# initialize library file from given arguments
library = genobox_modules.initialize_library(args.libfile, args.se, args.pe1, args.pe2, args.sample, args.mapq, args.libs, args.pl)
# start run
if args.sample:
print "--------------------------------------"
print "Processing sample: %s" % args.sample
print "--------------------------------------"
print "Starting alignment"
(bamfiles, library) = start_alignment(args, logger)
print "Starting bam processing"
final_bam = start_bamprocess(library, genobox_modules.unique(bamfiles.values()), args.mapq, args.libs, args.tmpdir, args.queue, final_bam, args.realignment, args.known, args.fa, args.sample, args.partition, logger)
# remove queuing system outfiles
genobox_modules.rm_files(['run_genobox_*', 'semaphores.*'])
print "Done"
print "--------------------------------------"
def start_alignment(args, logger):
'''Start alignment of fastq files using BWA'''
import genobox_modules
from genobox_classes import Semaphore, Library
import subprocess
import os
import random
import string
paths = genobox_modules.setSystem()
home = os.getcwd()
semaphore_ids = []
bamfiles = dict()
if not os.path.exists('alignment'):
os.makedirs('alignment')
# initialize library file from given arguments (if args.mapq is defined then its called from abgv, else it is called from alignment)
if hasattr(args, 'mapq'):
library = genobox_modules.initialize_library(args.libfile, args.se, args.pe1, args.pe2, args.sample, args.mapq, args.libs, args.pl)
else:
library = genobox_modules.initialize_library(args.libfile, args.se, args.pe1, args.pe2, args.sample, [30], args.libs, args.pl)
# check for fa
check_fa(args.fa, args.bwa6)
# check for if trimming was performed (abgv only) and set correct files
#(se_files, pe1_files, pe2_files) = check_trim(args)
# start single end alignments
if args.se:
# get platform info
(PL, PL2data) = library.getPL('Data')
print "Submitting single end alignments"
for key,value in PL2data.items():
if key == 'ILLUMINA' or key == 'HELICOS':
fqtypes_se = []
# filter to only contain single end files
toalign = []
for v in value:
if v in args.se: toalign.append(v)
for fq in toalign: fqtypes_se.append(check_formats_fq(fq, args.gz, args.bwa6))
# submit
(se_align_ids, bamfiles_se) = bwa_se_align(toalign, args.fa, fqtypes_se, args.qtrim, args.N, 'alignment/', args.bwa6, library, args.n, args.queue, args.add_aln, args.partition, logger)
semaphore_ids.extend(se_align_ids)
bamfiles.update(bamfiles_se)
elif key == 'PACBIO':
toalign = []
for v in value:
if v in args.se: toalign.append(v)
fqtypes_se = []
for fq in toalign: fqtypes_se.append(check_formats_fq(fq, args.gz, args.bwa6))
(se_align_ids, bamfiles_se) = bwasw_pacbio(toalign, args.fa, fqtypes_se, 'alignment/', args.bwa6, library, args.n, args.queue, args.partition, logger)
semaphore_ids.extend(se_align_ids)
bamfiles.update(bamfiles_se)
elif key == 'IONTORRENT':
toalign = []
for v in value:
if v in args.se: toalign.append(v)
fqtypes_se = []
for fq in toalign: fqtypes_se.append(check_formats_fq(fq, args.gz, args.bwa6))
(se_align_ids, bamfiles_se) = bwasw_iontorrent(toalign, args.fa, fqtypes_se, 'alignment/', args.bwa6, library, args.n, args.queue, args.partition, logger)
semaphore_ids.extend(se_align_ids)
bamfiles.update(bamfiles_se)
# start paired end alignments
if args.pe1:
if len(args.pe1) != len(args.pe2):
raise ValueError('Same number of files must be given to --pe1 and --pe2')
# set fqtypes
fqtypes_pe1 = []
fqtypes_pe2 = []
for fq in args.pe1: fqtypes_pe1.append(check_formats_fq(fq, args.gz, args.bwa6))
for fq in args.pe2: fqtypes_pe2.append(check_formats_fq(fq, args.gz, args.bwa6))
print "Submitting paired end alignments"
(pe_align_ids, bamfiles_pe) = bwa_pe_align(args.pe1, args.pe2, args.fa, fqtypes_pe1, fqtypes_pe2, args.qtrim, args.N, 'alignment/', args.bwa6, args.a, library, args.n, args.queue, args.add_aln, args.partition, logger)
semaphore_ids.extend(pe_align_ids)
bamfiles.update(bamfiles_pe)
# update library
library.update_with_tag('Data', 'BAM', bamfiles, True)
# wait for jobs to finish
print "Waiting for jobs to finish ..."
s = Semaphore(semaphore_ids, home, 'bwa_alignment', args.queue, 60, 172800)
s.wait()
print "--------------------------------------"
# return bamfiles
return (bamfiles, library)
def start_abgv(args, logger):
'''Start alignment, bam processing, genotyping, vcffiltering, dbsnp annotation, bcf2ref'''
import os
import subprocess
# check genome file
genobox_modules.check_genome(args.genome)
final_bam = 'alignment/%s.flt.sort.rmdup.bam' % args.sample
final_bcf = 'genotyping/%s.all.bcf' % args.sample
# initialize library file from given arguments
library = genobox_modules.initialize_library(args.libfile, args.se,
args.pe1, args.pe2,
args.sample, args.mapq,
args.libs, args.pl)
# start run
if args.sample:
print "--------------------------------------"
print "Processing sample: %s" % args.sample
print "--------------------------------------"
# toggle start trimming
#if args.no_trim == False:
# print "Starting trimming"
# (se_files, pe1_files, pe2_files) = start_trim(args, logger)
# library.update(Trim=se_files+pe1_files+pe2_files)
print "Starting alignment"
(bamfiles, library) = start_alignment(args, logger)
print "Starting bam processing"
final_bam = start_bamprocess(library,
genobox_modules.unique(bamfiles.values()),
args.mapq, args.libs, args.tmpdir, args.queue,
final_bam, args.realignment, args.known,
args.fa, args.sample, args.partition, logger)
print "Starting bam stats"
start_bamstats(args, final_bam, args.partition, logger, wait=False)
print "Starting genotyping"
if args.caller == 'samtools':
final_bcf = start_genotyping(final_bam, args.genome, args.fa,
args.prior, args.pp, args.queue,
final_bcf, args.sample, args.partition,
logger)
print "Starting vcffiltering"
final_vcf = start_vcffilter(final_bcf, args.genome, args.caller,
args.Q, args.ex, args.rmsk, args.ab,
args.prune, args.ovar, args.queue,
args.sample, args.partition, logger)
print "Start dbsnp"
final_dbsnp_vcf = start_dbsnp(final_vcf, args.ex, args.dbsnp,
args.ovar, args.queue, args.partition,
logger)
print "Start bcf2ref"
start_bcf2ref(final_bcf, args.genome, args.Q, args.ex, args.dbsnp,
args.rmsk, 'genotyping/indels_for_filtering.vcf',
args.oref, args.queue, args.sample, args.partition,
logger)
elif args.caller == 'gatk':
print "Start genotyping (gatk)"
vcffiles = start_genotyping_gatk(final_bam, args.genome, args.fa,
args.dbsnp, args.call_conf,
args.args.call_emit, args.output_mode,
args.queue, args.sample,
args.partition, logger)
print "Start vcffiltering (gatk)"
final_vcfs = start_vcffilter_gatk(vcffiles, args.genome, args.fa,
args.Q, args.rmsk, args.ab,
args.prune, args.queue, args.sample,
args.partition, args.logger)
# remove queuing system outfiles
genobox_modules.rm_files(['run_genobox_*', 'semaphores.*'])
print "Done"
print "--------------------------------------"
print "Raw genotyping is written in genotyping/all.bcf"
print "High confidence variants: %s" % args.ovar
print "High confidence reference: %s" % args.oref
print "--------------------------------------"
def start_abgv(args, logger):
"""Start alignment, bam processing, genotyping, vcffiltering, dbsnp annotation, bcf2ref"""
import os
import subprocess
# check genome file
genobox_modules.check_genome(args.genome)
final_bam = "alignment/%s.flt.sort.rmdup.bam" % args.sample
final_bcf = "genotyping/%s.all.bcf" % args.sample
# initialize library file from given arguments
library = genobox_modules.initialize_library(
args.libfile, args.se, args.pe1, args.pe2, args.sample, args.mapq, args.libs, args.pl
)
# start run
if args.sample:
print "--------------------------------------"
print "Processing sample: %s" % args.sample
print "--------------------------------------"
# toggle start trimming
# if args.no_trim == False:
# print "Starting trimming"
# (se_files, pe1_files, pe2_files) = start_trim(args, logger)
# library.update(Trim=se_files+pe1_files+pe2_files)
print "Starting alignment"
(bamfiles, library) = start_alignment(args, logger)
print "Starting bam processing"
final_bam = start_bamprocess(
library,
genobox_modules.unique(bamfiles.values()),
args.mapq,
args.libs,
args.tmpdir,
args.queue,
final_bam,
args.realignment,
args.known,
args.fa,
args.sample,
args.partition,
logger,
)
print "Starting bam stats"
start_bamstats(args, final_bam, args.partition, logger, wait=False)
print "Starting genotyping"
if args.caller == "samtools":
final_bcf = start_genotyping(
final_bam,
args.genome,
args.fa,
args.prior,
args.pp,
args.queue,
final_bcf,
args.sample,
args.partition,
logger,
)
print "Starting vcffiltering"
final_vcf = start_vcffilter(
final_bcf,
args.genome,
args.caller,
args.Q,
args.ex,
args.rmsk,
args.ab,
args.prune,
args.ovar,
args.queue,
args.sample,
args.partition,
logger,
)
print "Start dbsnp"
final_dbsnp_vcf = start_dbsnp(final_vcf, args.ex, args.dbsnp, args.ovar, args.queue, args.partition, logger)
print "Start bcf2ref"
start_bcf2ref(
final_bcf,
args.genome,
args.Q,
args.ex,
args.dbsnp,
args.rmsk,
"genotyping/indels_for_filtering.vcf",
args.oref,
args.queue,
args.sample,
args.partition,
logger,
)
elif args.caller == "gatk":
print "Start genotyping (gatk)"
vcffiles = start_genotyping_gatk(
final_bam,
args.genome,
args.fa,
args.dbsnp,
args.call_conf,
args.args.call_emit,
args.output_mode,
args.queue,
args.sample,
args.partition,
logger,
)
print "Start vcffiltering (gatk)"
final_vcfs = start_vcffilter_gatk(
vcffiles,
args.genome,
args.fa,
args.Q,
args.rmsk,
args.ab,
args.prune,
args.queue,
args.sample,
args.partition,
args.logger,
)
# remove queuing system outfiles
genobox_modules.rm_files(["run_genobox_*", "semaphores.*"])
print "Done"
print "--------------------------------------"
print "Raw genotyping is written in genotyping/all.bcf"
print "High confidence variants: %s" % args.ovar
print "High confidence reference: %s" % args.oref
print "--------------------------------------"
def start_bamprocess(library_file, bams, mapq, libs, tmpdir, queue, final_bam,
realignment, known, fa, sample, partition, logger):
'''Starts bam processing of input files'''
import subprocess
import genobox_modules
from genobox_classes import Moab, Semaphore, Library
import os
# set queueing
paths = genobox_modules.setSystem()
home = os.getcwd()
cpuA = 'nodes=1:ppn=1,mem=512mb,walltime=345600'
cpuC = 'nodes=1:ppn=1,mem=2gb,walltime=345600'
cpuE = 'nodes=1:ppn=1,mem=5gb,walltime=345600'
cpuF = 'nodes=1:ppn=2,mem=2gb,walltime=345600'
cpuB = 'nodes=1:ppn=16,mem=10gb,walltime=345600'
cpuG = 'nodes=1:ppn=1,mem=6gb,walltime=345600'
cpuH = 'nodes=1:ppn=2,mem=7gb,walltime=345600'
# create library instance
if library_file and library_file != 'None':
if isinstance(library_file, Library):
library = library_file
else:
library = Library(library_file)
library.read()
else:
library = genobox_modules.initialize_library(libfile=library_file,
sample=sample,
mapq=mapq,
libs=libs,
bams=bams)
(bam2lib, lib2bam) = library.getBamLibs()
## CREATE CALLS ##
# filter bam and sort
(filter_sort_calls,
filter_sort_files) = bam_filter_sort(lib2bam, bam2lib, 1500000000)
# merge to libs
(merge_lib_calls, librarys) = merge_bam(lib2bam.keys(),
lib2bam.values(),
add_suffix=True,
final_suffix='.flt.sort.bam',
tmpdir=tmpdir)
# rmdup on libs
(rmdup_calls, rmdup_files) = rmdup(librarys, tmpdir)
# optional: realignment
if realignment:
(merge_final_call, sample_file) = merge_bam([final_bam], [rmdup_files],
add_suffix=False)
(realign_calls, final_file) = realign_bam(final_bam, final_bam, fa,
known)
else:
# merge to final file
(merge_final_call, final_file) = merge_bam([final_bam], [rmdup_files],
add_suffix=False)
## SUBMIT JOBS ##
print "Submitting jobs"
filtersort_moab = Moab(filter_sort_calls,
logfile=logger,
runname='run_genobox_filtersort',
queue=queue,
cpu=cpuH,
partition=partition)
mergelib_moab = Moab(merge_lib_calls,
logfile=logger,
runname='run_genobox_lib_merge',
queue=queue,
cpu=cpuE,
depend=True,
depend_type='complex',
depend_val=map(len, lib2bam.values()),
depend_ids=filtersort_moab.ids,
partition=partition)
rmdup_moab = Moab(
rmdup_calls,
logfile=logger,
runname='run_genobox_rmdup',
queue=queue,
cpu=cpuG,
depend=True,
depend_type='one2one',
depend_val=[1],
depend_ids=mergelib_moab.ids,
partition=partition
) # NB: If memory should be changed, also change java memory spec in rmdup function
mergefinal_moab = Moab(merge_final_call,
logfile=logger,
runname='run_genobox_final_merge',
queue=queue,
cpu=cpuC,
depend=True,
depend_type='conc',
depend_val=[len(rmdup_moab.ids)],
depend_ids=rmdup_moab.ids,
partition=partition)
if realignment:
realign_moab = Moab(realign_calls,
logfile=logger,
runname='run_genobox_realignment',
queue=queue,
cpu=cpuE,
depend=True,
depend_type='one2one',
depend_val=[1],
depend_ids=mergefinal_moab.ids,
partition=partition)
# realignment calls needs to be written together in a shell-file or dependent on each other #
# release jobs #
print "Releasing jobs"
#filtersort_moab.release()
#mergelib_moab.release()
#rmdup_moab.release()
#mergefinal_moab.release()
#if realignment: realign_moab.release()
# semaphore
print "Waiting for jobs to finish ..."
if realignment:
s = Semaphore(realign_moab.ids, home, 'bam_processing', queue, 20,
345600)
else:
s = Semaphore(mergefinal_moab.ids, home, 'bam_processing', queue, 20,
345600)
s.wait()
print "--------------------------------------"
# return final bamfile
return final_bam
def start_bamprocess(library_file, bams, mapq, libs, tmpdir, queue, final_bam, realignment, known, fa, sample, partition, logger):
'''Starts bam processing of input files'''
import subprocess
import genobox_modules
from genobox_classes import Moab, Semaphore, Library
import os
# set queueing
paths = genobox_modules.setSystem()
home = os.getcwd()
cpuA = 'nodes=1:ppn=1,mem=512mb,walltime=345600'
cpuC = 'nodes=1:ppn=1,mem=2gb,walltime=345600'
cpuE = 'nodes=1:ppn=1,mem=5gb,walltime=345600'
cpuF = 'nodes=1:ppn=2,mem=2gb,walltime=345600'
cpuB = 'nodes=1:ppn=16,mem=10gb,walltime=345600'
cpuH = 'nodes=1:ppn=2,mem=7gb,walltime=345600'
# create library instance
if library_file and library_file != 'None':
if isinstance(library_file, Library):
library = library_file
else:
library = Library(library_file)
library.read()
else:
library = genobox_modules.initialize_library(libfile=library_file, sample=sample, mapq=mapq, libs=libs, bams=bams)
(bam2lib, lib2bam) = library.getBamLibs()
## CREATE CALLS ##
# filter bam and sort
(filter_sort_calls, filter_sort_files) = bam_filter_sort(lib2bam, bam2lib, 1500000000)
# merge to libs
(merge_lib_calls, librarys) = merge_bam(lib2bam.keys(), lib2bam.values(), add_suffix=True, final_suffix='.flt.sort.bam', tmpdir=tmpdir)
# rmdup on libs
(rmdup_calls, rmdup_files) = rmdup(librarys, tmpdir)
# optional: realignment
if realignment:
(merge_final_call, sample_file) = merge_bam([final_bam], [rmdup_files], add_suffix=False)
(realign_calls, final_file) = realign_bam(final_bam, final_bam, fa, known)
else:
# merge to final file
(merge_final_call, final_file) = merge_bam([final_bam], [rmdup_files], add_suffix=False)
## SUBMIT JOBS ##
print "Submitting jobs"
filtersort_moab = Moab(filter_sort_calls, logfile=logger, runname='run_genobox_filtersort', queue=queue, cpu=cpuH, partition=partition)
mergelib_moab = Moab(merge_lib_calls, logfile=logger, runname='run_genobox_lib_merge', queue=queue, cpu=cpuE, depend=True, depend_type='complex', depend_val=map(len, lib2bam.values()), depend_ids=filtersort_moab.ids, partition=partition)
rmdup_moab = Moab(rmdup_calls, logfile=logger, runname='run_genobox_rmdup', queue=queue, cpu=cpuE, depend=True, depend_type='one2one', depend_val=[1], depend_ids=mergelib_moab.ids, partition=partition) # NB: If memory should be changed, also change java memory spec in rmdup function
mergefinal_moab = Moab(merge_final_call, logfile=logger, runname='run_genobox_final_merge', queue=queue, cpu=cpuC, depend=True, depend_type='conc', depend_val=[len(rmdup_moab.ids)], depend_ids=rmdup_moab.ids, partition=partition)
if realignment:
realign_moab = Moab(realign_calls, logfile=logger, runname='run_genobox_realignment', queue=queue, cpu=cpuE, depend=True, depend_type='one2one', depend_val=[1], depend_ids=mergefinal_moab.ids, partition=partition)
# realignment calls needs to be written together in a shell-file or dependent on each other #
# release jobs #
print "Releasing jobs"
#filtersort_moab.release()
#mergelib_moab.release()
#rmdup_moab.release()
#mergefinal_moab.release()
#if realignment: realign_moab.release()
# semaphore
print "Waiting for jobs to finish ..."
if realignment:
s = Semaphore(realign_moab.ids, home, 'bam_processing', queue, 20, 2*86400)
else:
s = Semaphore(mergefinal_moab.ids, home, 'bam_processing', queue, 20, 2*86400)
s.wait()
print "--------------------------------------"
# return final bamfile
return final_bam
def start_alignment(args, logger):
'''Start alignment of fastq files using BWA'''
import genobox_modules
from genobox_classes import Semaphore, Library
import subprocess
import os
import random
import string
paths = genobox_modules.setSystem()
home = os.getcwd()
semaphore_ids = []
bamfiles = dict()
if not os.path.exists('alignment'):
os.makedirs('alignment')
# initialize library file from given arguments (if args.mapq is defined then its called from abgv, else it is called from alignment)
if hasattr(args, 'mapq'):
library = genobox_modules.initialize_library(args.libfile, args.se,
args.pe1, args.pe2,
args.sample, args.mapq,
args.libs, args.pl)
else:
library = genobox_modules.initialize_library(args.libfile, args.se,
args.pe1, args.pe2,
args.sample, [30],
args.libs, args.pl)
# check for fa
check_fa(args.fa, args.bwa6)
# check for if trimming was performed (abgv only) and set correct files
#(se_files, pe1_files, pe2_files) = check_trim(args)
# start single end alignments
if args.se:
# get platform info
(PL, PL2data) = library.getPL('Data')
print "Submitting single end alignments"
for key, value in PL2data.items():
if key == 'ILLUMINA' or key == 'HELICOS':
fqtypes_se = []
# filter to only contain single end files
toalign = []
for v in value:
if v in args.se: toalign.append(v)
for fq in toalign:
if args.quals:
fqtypes_se.append(args.quals)
else:
fqtypes_se.append(
check_formats_fq(fq, args.gz, args.bwa6))
# submit
(se_align_ids, bamfiles_se) = bwa_se_align(
toalign, args.fa, fqtypes_se, args.qtrim, args.N,
'alignment/', args.bwa6, library, args.n, args.queue,
args.add_aln, args.partition, logger)
semaphore_ids.extend(se_align_ids)
bamfiles.update(bamfiles_se)
elif key == 'PACBIO':
toalign = []
for v in value:
if v in args.se: toalign.append(v)
fqtypes_se = []
for fq in toalign:
if args.quals:
fqtypes_se.append(args.quals)
else:
fqtypes_se.append(
check_formats_fq(fq, args.gz, args.bwa6))
# submit
(se_align_ids, bamfiles_se) = bwasw_pacbio(
toalign, args.fa, fqtypes_se, 'alignment/', args.bwa6,
library, args.n, args.queue, args.partition, logger)
semaphore_ids.extend(se_align_ids)
bamfiles.update(bamfiles_se)
elif key == 'IONTORRENT' or key == '454':
toalign = []
for v in value:
if v in args.se: toalign.append(v)
fqtypes_se = []
for fq in toalign:
if args.quals:
fqtypes_se.append(args.quals)
else:
fqtypes_se.append(
check_formats_fq(fq, args.gz, args.bwa6))
# submit
(se_align_ids, bamfiles_se) = bwasw_iontorrent(
toalign, args.fa, fqtypes_se, 'alignment/', args.bwa6,
library, args.n, args.queue, args.partition, logger)
semaphore_ids.extend(se_align_ids)
bamfiles.update(bamfiles_se)
# start paired end alignments
if args.pe1:
if len(args.pe1) != len(args.pe2):
raise ValueError(
'Same number of files must be given to --pe1 and --pe2')
# set fqtypes
fqtypes_pe1 = []
fqtypes_pe2 = []
for fq in args.pe1:
if args.quals:
fqtypes_pe1.append(args.quals)
else:
fqtypes_pe1.append(check_formats_fq(fq, args.gz, args.bwa6))
for fq in args.pe2:
if args.quals:
fqtypes_pe2.append(args.quals)
else:
fqtypes_pe2.append(check_formats_fq(fq, args.gz, args.bwa6))
# submit
print "Submitting paired end alignments"
(pe_align_ids,
bamfiles_pe) = bwa_pe_align(args.pe1, args.pe2, args.fa, fqtypes_pe1,
fqtypes_pe2, args.qtrim, args.N,
'alignment/', args.bwa6, args.a, library,
args.n, args.queue, args.add_aln,
args.partition, logger)
semaphore_ids.extend(pe_align_ids)
bamfiles.update(bamfiles_pe)
# update library
library.update_with_tag('Data', 'BAM', bamfiles, True)
# wait for jobs to finish
print "Waiting for jobs to finish ..."
s = Semaphore(semaphore_ids, home, 'bwa_alignment', args.queue, 60, 345600)
s.wait()
print "--------------------------------------"
# return bamfiles
return (bamfiles, library)