def makeAggregate(cells, directory, suffix, output):
"""
Create aggregate sample.
Make an aggregate bam file from a list of cells, sorts and indexes
the file for easy use in IGV. Suffix is required to prevent non
0-padded numbers matching the wrong files. Return final file name.
Parameters
----------
cells : list
List of cell names to create aggregate from.
directory : string
Directory path with the bam files from each cell.
suffix : string
String to match the end of the bam file, use to add file extension
and to anchor the extension after file numbers - this will prevent
cell_4 matching cell_4*.
output : string
String containing output file location.
"""
from glob import glob
cells = set(cells)
fileList = []
for cell in cells:
fileList.append(glob(os.path.join(directory, "*" + cell + suffix))[0])
pysam.cat("-o", output + ".bam", *fileList, catch_stdout=False)
pysam.sort(output + ".bam", output + ".sorted", catch_stdout=False)
pysam.index(output + ".sorted.bam", catch_stdout=False)
return output + ".sorted.bam"
def makeAggregate(cells, directory, suffix, output):
"""
Create aggregate sample.
Make an aggregate bam file from a list of cells, sorts and indexes
the file for easy use in IGV. Suffix is required to prevent non
0-padded numbers matching the wrong files. Return final file name.
Parameters
----------
cells : list
List of cell names to create aggregate from.
directory : string
Directory path with the bam files from each cell.
suffix : string
String to match the end of the bam file, use to add file extension
and to anchor the extension after file numbers - this will prevent
cell_4 matching cell_4*.
output : string
String containing output file location.
"""
from glob import glob
cells = set(cells)
fileList = []
for cell in cells:
fileList.append(glob(os.path.join(directory, "*" + cell + suffix))[0])
pysam.cat("-o", output + ".bam", *fileList, catch_stdout=False)
pysam.sort("-T", output + "_tmp", output + ".bam", "-o", output + ".sorted.bam", catch_stdout=False)
pysam.index(output + ".sorted.bam", catch_stdout=False)
return output + ".sorted.bam"
def collect_bam_chunks(inpath, chrs, outpath, unmapped):
allpaths = [inpath + ".tmp." + c + ".bam" for c in chrs]
if unmapped:
allpaths = [inpath + ".tmp." + c + ".bam" for c in chrs[:-1]]
allpaths.append(inpath + ".tmp." + "unmapped" + ".bam")
cat_args = ['-o', outpath] + allpaths
pysam.cat(*cat_args)
x = [os.remove(f) for f in allpaths]
def merge_bam(bam_collection,
output_path,
template_bam=None,
sort_order=None,
threads=1):
"""Merge a readname sorted collection of BAM files."""
bam_collection = [bam for bam in bam_collection if os.path.exists(bam)]
if not template_bam:
template_bam = bam_collection[0]
pysam.cat('-h', template_bam, '-o', output_path, *bam_collection)
for bam in bam_collection:
try:
os.remove(bam)
except OSError:
pass
if sort_order:
sort_bam(inpath=output_path,
output=output_path,
sort_order=sort_order,
threads=threads)
return output_path
def run_merge_types():
bam_paths = sys.argv[1:]
dum_obj = pysam.Samfile(sys.argv[1],"rb")
unique = int(time.time())
multi_path = "merge_multi_%d.bam" % (unique,)
multi_out = pysam.Samfile(multi_path,"wb",template=dum_obj)
start = time.time()
print "Starting Multi File Merge"
for cur_path in bam_paths:
cur_obj = pysam.Samfile(cur_path,"rb")
for alig in cur_obj.fetch(until_eof=True):
multi_out.write(alig)
cur_obj.close()
multi_out.close()
print "Multi took %d" % (int(time.time() - start))
single_path = "merge_single_%d.bam" % (unique,)
single_out = pysam.Samfile(single_path,"wb",template=dum_obj)
start = time.time()
single_obj = pysam.Samfile(multi_path,"rb")
print "Starting Single File Merge"
for alig in single_obj.fetch(until_eof=True):
single_out.write(alig)
print "Single took %d" % (int(time.time() - start))
os.remove(multi_path)
os.remove(single_path)
print "Starting Samtools Cat"
start = time.time()
cat_path = "merge_cat_%d.bam" % (unique,)
pysam.cat("-o",cat_path,*bam_paths)
print "Samtools took %d" % (int(time.time() - start))
dum_obj.close()
os.remove(cat_path)
def premap_stampy(data_folder,
adaID,
VERBOSE=0,
threads=1,
summary=True,
maxreads=-1,
subsrate=0.05,
gapopen=40,
gapextend=3):
'''Call stampy for actual mapping'''
if VERBOSE:
print 'Premapping: adaID ', adaID
if summary:
summary_filename = get_premap_summary_filename(data_folder, adaID)
# Stampy can handle both gzipped and uncompressed fastq inputs
input_filenames = get_read_filenames(data_folder, adaID, gzip=True)
if not os.path.isfile(input_filenames[0]):
input_filenames = get_read_filenames(data_folder, adaID, gzip=False)
if not all(map(os.path.isfile, input_filenames)):
raise OSError('Input files for mapping not found: ' +
input_filenames[0])
# parallelize if requested
if threads == 1:
call_list = [
stampy_bin,
'--overwrite',
'-g',
get_reference_premap_index_filename(data_folder, adaID, ext=False),
'-h',
get_reference_premap_hash_filename(data_folder, adaID, ext=False),
'-o',
get_premapped_filename(data_folder, adaID, type='sam'),
'--insertsize=450',
'--insertsd=100',
'--substitutionrate=' + str(subsrate),
'--gapopen=' + str(gapopen),
'--gapextend=' + str(gapextend),
]
if maxreads > 0:
call_list.append('--numrecords=' + str(maxreads))
call_list.extend(['-M'] + input_filenames)
call_list = map(str, call_list)
if VERBOSE >= 2:
print ' '.join(call_list)
sp.call(call_list)
if summary:
with open(get_premap_summary_filename(data_folder, adaID),
'a') as f:
f.write('\nStampy premapped (single thread).\n')
# Convert to compressed BAM
convert_sam_to_bam(
get_premapped_filename(data_folder, adaID, type='bam'))
if summary:
with open(summary_filename, 'a') as f:
f.write('\nSAM file converted to compressed BAM: '+\
get_premapped_filename(data_folder, adaID, type='bam')+'\n')
else:
# Multithreading works as follows: call qsub + stampy, monitor the process
# IDs with qstat at regular intervals, and finally merge results with pysam
output_file_parts = [
get_premapped_filename(data_folder,
adaID,
type='bam',
part=(j + 1)) for j in xrange(threads)
]
# Submit map script
jobs_done = np.zeros(threads, bool)
job_IDs = np.zeros(threads, 'S30')
# Submit map call
import hivwholeseq
JOBDIR = hivwholeseq.__path__[0].rstrip('/') + '/'
JOBLOGOUT = JOBDIR + 'logout'
JOBLOGERR = JOBDIR + 'logerr'
cluster_time = ['23:59:59', '1:59:59']
vmem = '8G'
for j in xrange(threads):
call_list = [
'qsub', '-cwd', '-b', 'y', '-S', '/bin/bash', '-o', JOBLOGOUT,
'-e', JOBLOGERR, '-N', adaID + ' p' + str(j + 1), '-l',
'h_rt=' + cluster_time[threads >= 30], '-l', 'h_vmem=' + vmem,
stampy_bin, '--overwrite', '-g',
get_reference_premap_index_filename(
data_folder, adaID, ext=False), '-h',
get_reference_premap_hash_filename(
data_folder, adaID, ext=False), '-o',
get_premapped_filename(
data_folder, adaID, type='sam', part=(j + 1)),
'--processpart=' + str(j + 1) + '/' + str(threads),
'--insertsize=450', '--insertsd=100', '--substitutionrate=' +
str(subsrate), '--gapopen=' + str(gapopen),
'--gapextend=' + str(gapextend), '-M'
] + input_filenames
call_list = map(str, call_list)
if VERBOSE >= 2:
print ' '.join(call_list)
job_ID = sp.check_output(call_list)
job_ID = job_ID.split()[2]
job_IDs[j] = job_ID
# Monitor output
time_wait = 10 # secs
while not jobs_done.all():
# Sleep some time
time.sleep(time_wait)
# Get the output of qstat to check the status of jobs
qstat_output = sp.check_output(['qstat'])
qstat_output = qstat_output.split(
'\n')[:-1] # The last is an empty line
if VERBOSE >= 3:
print qstat_output
if len(qstat_output) < 3:
jobs_done[:] = True
break
else:
qstat_output = [line.split()[0] for line in qstat_output[2:]]
time_wait = 10 # secs
for j in xrange(threads):
if jobs_done[j]:
continue
if job_IDs[j] not in qstat_output:
# Convert to BAM for merging
if VERBOSE >= 1:
print 'Convert premapped reads to BAM for merging: adaID '+\
adaID+', part '+str(j+1)+ ' of '+ \
str(threads)
convert_sam_to_bam(output_file_parts[j])
# We do not need to wait if we did the conversion (it takes
# longer than some secs)
time_wait = 0
jobs_done[j] = True
if summary:
with open(summary_filename, 'a') as f:
f.write('Stampy premapped (' + str(threads) + ' threads).\n')
# Concatenate output files
if VERBOSE >= 1:
print 'Concatenate premapped reads: adaID ' + adaID + '...',
output_filename = get_premapped_filename(data_folder,
adaID,
type='bam',
unsorted=True)
pysam.cat('-o', output_filename, *output_file_parts)
if VERBOSE >= 1:
print 'done.'
if summary:
with open(summary_filename, 'a') as f:
f.write('BAM files concatenated (unsorted).\n')
# Sort the file by read names (to ensure the pair_generator)
# NOTE: we exclude the extension and the option -f because of a bug in samtools
if VERBOSE >= 1:
print 'Sort premapped reads: adaID ' + adaID
output_filename_sorted = get_premapped_filename(data_folder,
adaID,
type='bam',
unsorted=False)
pysam.sort('-n', output_filename, output_filename_sorted[:-4])
if summary:
with open(summary_filename, 'a') as f:
f.write('Joint BAM file sorted.\n')
# Reheader the file without BAM -> SAM -> BAM
if VERBOSE >= 1:
print 'Reheader premapped reads: adaID ' + adaID
header_filename = get_premapped_filename(data_folder,
adaID,
type='sam',
part=1)
pysam.reheader(header_filename, output_filename_sorted)
if summary:
with open(summary_filename, 'a') as f:
f.write('Joint BAM file reheaded.\n')
if VERBOSE >= 1:
print 'Remove temporary files: adaID ' + adaID
remove_premapped_tempfiles(data_folder, adaID, VERBOSE=VERBOSE)
if summary:
with open(summary_filename, 'a') as f:
f.write('Temp premapping files removed.\n')
f.write('\n')
def map_stampy(data_folder, adaID, fragment, VERBOSE=0, threads=1,
cluster_time='23:59:59', maxreads=-1, summary=True,
rescue=False, dry=False):
'''Map using stampy'''
frag_gen = fragment[:2]
if summary:
summary_filename = get_map_summary_filename(data_folder, adaID, frag_gen,
rescue=rescue)
# Set mapping penalty scores: softer for rescues and F3 and F5
global subsrate
if rescue:
subsrate = '0.2'
stampy_gapopen = 5 # Default: 40
stampy_gapextend = 1 # Default: 3
elif frag_gen not in ('F3', 'F5'):
stampy_gapopen = 60 # Default: 40
stampy_gapextend = 5 # Default: 3
else:
stampy_gapopen = 30 # Default: 40
stampy_gapextend = 2 # Default: 3
if VERBOSE:
print 'Map via stampy: '+adaID+' '+frag_gen
if not rescue:
input_filename = get_divided_filename(data_folder, adaID, fragment, type='bam')
# NOTE: we introduced fragment nomenclature late, e.g. F3a. Check for that
if not os.path.isfile(input_filename):
if frag_gen == 'F3':
input_filename = input_filename.replace('F3a', 'F3')
else:
input_filename = get_divided_filename(data_folder, adaID, 'unmapped', type='bam')
# Check existance of input file, because stampy creates output anyway
if not os.path.isfile(input_filename):
if summary:
with open(summary_filename, 'a') as f:
f.write('Failed (input file for mapping not found).\n')
raise ValueError(samplename+', fragment '+fragment+': input file not found.')
# parallelize if requested
if threads == 1:
output_filename = get_mapped_filename(data_folder, adaID, frag_gen, type='sam',
rescue=rescue)
# Map
call_list = [stampy_bin,
'-g', get_index_file(data_folder, adaID, frag_gen, ext=False),
'-h', get_hash_file(data_folder, adaID, frag_gen, ext=False),
'-o', output_filename,
'--overwrite',
'--substitutionrate='+subsrate,
'--gapopen', stampy_gapopen,
'--gapextend', stampy_gapextend]
if stampy_sensitive:
call_list.append('--sensitive')
# Take only a (random) subsample: stampy uses the fraction of reads
# intead of the number
if maxreads > 0:
# FIXME: figure out the -s option and the --numrecords option
call_list.extend(['--numrecords', maxreads])
#n_pairs_tot = get_number_reads(input_filename, 'bam') / 2
#frac_pairs = 1.0 * maxreads / n_pairs_tot
#random_seed = np.random.randint(1e5)
#call_list.extend(['-s', frac_pairs + random_seed])
call_list = call_list + ['-M', input_filename]
call_list = map(str, call_list)
if VERBOSE >=2:
print ' '.join(call_list)
if not dry:
sp.call(call_list)
if summary:
with open(summary_filename, 'a') as f:
f.write('Stampy mapped (single thread).\n')
output_filename = get_mapped_filename(data_folder, adaID, frag_gen, type='bam',
rescue=rescue)
convert_sam_to_bam(output_filename)
else:
if summary:
with open(summary_filename, 'a') as f:
f.write('Dry run works (single thread).\n')
if VERBOSE >= 1:
print 'Dry run works (single thread)'
return
else:
# Submit map script
jobs_done = np.zeros(threads, bool)
job_IDs = np.zeros(threads, 'S30')
for j in xrange(threads):
# Get output filename
output_filename = get_mapped_filename(data_folder, adaID, frag_gen,
type='sam', part=(j+1), rescue=rescue)
# Map
call_list = ['qsub','-cwd',
'-b', 'y',
'-S', '/bin/bash',
'-o', JOBLOGOUT,
'-e', JOBLOGERR,
'-N', 'm'+adaID.replace('-', '')+frag_gen+str(j+1),
'-l', 'h_rt='+cluster_time,
'-l', 'h_vmem='+vmem,
stampy_bin,
'-g', get_index_file(data_folder, adaID, frag_gen, ext=False),
'-h', get_hash_file(data_folder, adaID, frag_gen, ext=False),
'-o', output_filename,
'--overwrite',
'--processpart='+str(j+1)+'/'+str(threads),
'--substitutionrate='+subsrate,
'--gapopen', stampy_gapopen,
'--gapextend', stampy_gapextend]
if stampy_sensitive:
call_list.append('--sensitive')
call_list = call_list + ['-M', input_filename]
call_list = map(str, call_list)
if VERBOSE >= 2:
print ' '.join(call_list)
if not dry:
job_ID = sp.check_output(call_list)
job_ID = job_ID.split()[2]
job_IDs[j] = job_ID
if dry:
if summary:
with open(summary_filename, 'a') as f:
f.write('Dry run works (multi thread).\n')
if VERBOSE >= 1:
print 'Dry run works (multi thread)'
return
# Monitor output
output_file_parts = [get_mapped_filename(data_folder, adaID, frag_gen,
type='bam', part=(j+1), rescue=rescue)
for j in xrange(threads)]
time_wait = 10 # secs
while not jobs_done.all():
# Sleep some time
time.sleep(time_wait)
# Get the output of qstat to check the status of jobs
qstat_output = sp.check_output(['qstat'])
qstat_output = qstat_output.split('\n')[:-1] # The last is an empty line
if len(qstat_output) < 3:
jobs_done[:] = True
break
else:
qstat_output = [line.split()[0] for line in qstat_output[2:]]
time_wait = 10 # secs
for j in xrange(threads):
if jobs_done[j]:
continue
if job_IDs[j] not in qstat_output:
# Convert to BAM for merging
if VERBOSE >= 1:
print 'Convert mapped reads to BAM for merging: adaID '+\
adaID+', fragment '+frag_gen+', part '+str(j+1)+ ' of '+ \
str(threads)
convert_sam_to_bam(output_file_parts[j])
# We do not need to wait if we did the conversion (it takes
# longer than some secs)
time_wait = 0
jobs_done[j] = True
if summary:
with open(summary_filename, 'a') as f:
f.write('Stampy mapped ('+str(threads)+' threads).\n')
# Concatenate output files
output_filename = get_mapped_filename(data_folder, adaID, frag_gen, type='bam',
unsorted=True, rescue=rescue)
if VERBOSE >= 1:
print 'Concatenate mapped reads: adaID '+adaID+', fragment '+frag_gen
pysam.cat('-o', output_filename, *output_file_parts)
if summary:
with open(summary_filename, 'a') as f:
f.write('BAM files concatenated (unsorted).\n')
# Sort the file by read names (to ensure the pair_generator)
output_filename_sorted = get_mapped_filename(data_folder, adaID, frag_gen,
type='bam',
unsorted=False,
rescue=rescue)
# NOTE: we exclude the extension and the option -f because of a bug in samtools
if VERBOSE >= 1:
print 'Sort mapped reads: adaID '+adaID+', fragment '+frag_gen
pysam.sort('-n', output_filename, output_filename_sorted[:-4])
if summary:
with open(summary_filename, 'a') as f:
f.write('Joint BAM file sorted.\n')
# Reheader the file without BAM -> SAM -> BAM
if VERBOSE >= 1:
print 'Reheader mapped reads: adaID '+adaID+', fragment '+frag_gen
header_filename = get_mapped_filename(data_folder, adaID, frag_gen,
type='sam', part=1, rescue=rescue)
pysam.reheader(header_filename, output_filename_sorted)
if summary:
with open(summary_filename, 'a') as f:
f.write('Joint BAM file reheaded.\n')
# FIXME: check whether temp files are all deleted
if VERBOSE >= 1:
print 'Remove temporary files: adaID '+adaID+', fragment '+frag_gen
remove_mapped_tempfiles(data_folder, adaID, frag_gen, VERBOSE=VERBOSE, rescue=rescue)
if summary:
with open(summary_filename, 'a') as f:
f.write('Temp mapping files removed.\n')
f.write('\n')
def map_stampy(data_folder,
adaID,
fragment,
VERBOSE=0,
threads=1,
cluster_time='23:59:59',
maxreads=-1,
summary=True,
rescue=False,
dry=False):
'''Map using stampy'''
frag_gen = fragment[:2]
if summary:
summary_filename = get_map_summary_filename(data_folder,
adaID,
frag_gen,
rescue=rescue)
# Set mapping penalty scores: softer for rescues and F3 and F5
global subsrate
if rescue:
subsrate = '0.2'
stampy_gapopen = 5 # Default: 40
stampy_gapextend = 1 # Default: 3
elif frag_gen not in ('F3', 'F5'):
stampy_gapopen = 60 # Default: 40
stampy_gapextend = 5 # Default: 3
else:
stampy_gapopen = 30 # Default: 40
stampy_gapextend = 2 # Default: 3
if VERBOSE:
print 'Map via stampy: ' + adaID + ' ' + frag_gen
if not rescue:
input_filename = get_divided_filename(data_folder,
adaID,
fragment,
type='bam')
# NOTE: we introduced fragment nomenclature late, e.g. F3a. Check for that
if not os.path.isfile(input_filename):
if frag_gen == 'F3':
input_filename = input_filename.replace('F3a', 'F3')
else:
input_filename = get_divided_filename(data_folder,
adaID,
'unmapped',
type='bam')
# Check existance of input file, because stampy creates output anyway
if not os.path.isfile(input_filename):
if summary:
with open(summary_filename, 'a') as f:
f.write('Failed (input file for mapping not found).\n')
raise ValueError(samplename + ', fragment ' + fragment +
': input file not found.')
# parallelize if requested
if threads == 1:
output_filename = get_mapped_filename(data_folder,
adaID,
frag_gen,
type='sam',
rescue=rescue)
# Map
call_list = [
stampy_bin, '-g',
get_index_file(data_folder, adaID, frag_gen, ext=False), '-h',
get_hash_file(data_folder, adaID, frag_gen, ext=False), '-o',
output_filename, '--overwrite', '--substitutionrate=' + subsrate,
'--gapopen', stampy_gapopen, '--gapextend', stampy_gapextend
]
if stampy_sensitive:
call_list.append('--sensitive')
# Take only a (random) subsample: stampy uses the fraction of reads
# intead of the number
if maxreads > 0:
# FIXME: figure out the -s option and the --numrecords option
call_list.extend(['--numrecords', maxreads])
#n_pairs_tot = get_number_reads(input_filename, 'bam') / 2
#frac_pairs = 1.0 * maxreads / n_pairs_tot
#random_seed = np.random.randint(1e5)
#call_list.extend(['-s', frac_pairs + random_seed])
call_list = call_list + ['-M', input_filename]
call_list = map(str, call_list)
if VERBOSE >= 2:
print ' '.join(call_list)
if not dry:
sp.call(call_list)
if summary:
with open(summary_filename, 'a') as f:
f.write('Stampy mapped (single thread).\n')
output_filename = get_mapped_filename(data_folder,
adaID,
frag_gen,
type='bam',
rescue=rescue)
convert_sam_to_bam(output_filename)
else:
if summary:
with open(summary_filename, 'a') as f:
f.write('Dry run works (single thread).\n')
if VERBOSE >= 1:
print 'Dry run works (single thread)'
return
else:
# Submit map script
jobs_done = np.zeros(threads, bool)
job_IDs = np.zeros(threads, 'S30')
for j in xrange(threads):
# Get output filename
output_filename = get_mapped_filename(data_folder,
adaID,
frag_gen,
type='sam',
part=(j + 1),
rescue=rescue)
# Map
call_list = [
'qsub', '-cwd', '-b', 'y', '-S', '/bin/bash', '-o', JOBLOGOUT,
'-e', JOBLOGERR, '-N',
'm' + adaID.replace('-', '') + frag_gen + str(j + 1), '-l',
'h_rt=' + cluster_time, '-l', 'h_vmem=' + vmem, stampy_bin,
'-g',
get_index_file(data_folder, adaID, frag_gen, ext=False), '-h',
get_hash_file(data_folder, adaID, frag_gen,
ext=False), '-o', output_filename, '--overwrite',
'--processpart=' + str(j + 1) + '/' + str(threads),
'--substitutionrate=' + subsrate, '--gapopen', stampy_gapopen,
'--gapextend', stampy_gapextend
]
if stampy_sensitive:
call_list.append('--sensitive')
call_list = call_list + ['-M', input_filename]
call_list = map(str, call_list)
if VERBOSE >= 2:
print ' '.join(call_list)
if not dry:
job_ID = sp.check_output(call_list)
job_ID = job_ID.split()[2]
job_IDs[j] = job_ID
if dry:
if summary:
with open(summary_filename, 'a') as f:
f.write('Dry run works (multi thread).\n')
if VERBOSE >= 1:
print 'Dry run works (multi thread)'
return
# Monitor output
output_file_parts = [
get_mapped_filename(data_folder,
adaID,
frag_gen,
type='bam',
part=(j + 1),
rescue=rescue) for j in xrange(threads)
]
time_wait = 10 # secs
while not jobs_done.all():
# Sleep some time
time.sleep(time_wait)
# Get the output of qstat to check the status of jobs
qstat_output = sp.check_output(['qstat'])
qstat_output = qstat_output.split(
'\n')[:-1] # The last is an empty line
if len(qstat_output) < 3:
jobs_done[:] = True
break
else:
qstat_output = [line.split()[0] for line in qstat_output[2:]]
time_wait = 10 # secs
for j in xrange(threads):
if jobs_done[j]:
continue
if job_IDs[j] not in qstat_output:
# Convert to BAM for merging
if VERBOSE >= 1:
print 'Convert mapped reads to BAM for merging: adaID '+\
adaID+', fragment '+frag_gen+', part '+str(j+1)+ ' of '+ \
str(threads)
convert_sam_to_bam(output_file_parts[j])
# We do not need to wait if we did the conversion (it takes
# longer than some secs)
time_wait = 0
jobs_done[j] = True
if summary:
with open(summary_filename, 'a') as f:
f.write('Stampy mapped (' + str(threads) + ' threads).\n')
# Concatenate output files
output_filename = get_mapped_filename(data_folder,
adaID,
frag_gen,
type='bam',
unsorted=True,
rescue=rescue)
if VERBOSE >= 1:
print 'Concatenate mapped reads: adaID ' + adaID + ', fragment ' + frag_gen
pysam.cat('-o', output_filename, *output_file_parts)
if summary:
with open(summary_filename, 'a') as f:
f.write('BAM files concatenated (unsorted).\n')
# Sort the file by read names (to ensure the pair_generator)
output_filename_sorted = get_mapped_filename(data_folder,
adaID,
frag_gen,
type='bam',
unsorted=False,
rescue=rescue)
# NOTE: we exclude the extension and the option -f because of a bug in samtools
if VERBOSE >= 1:
print 'Sort mapped reads: adaID ' + adaID + ', fragment ' + frag_gen
pysam.sort('-n', output_filename, output_filename_sorted[:-4])
if summary:
with open(summary_filename, 'a') as f:
f.write('Joint BAM file sorted.\n')
# Reheader the file without BAM -> SAM -> BAM
if VERBOSE >= 1:
print 'Reheader mapped reads: adaID ' + adaID + ', fragment ' + frag_gen
header_filename = get_mapped_filename(data_folder,
adaID,
frag_gen,
type='sam',
part=1,
rescue=rescue)
pysam.reheader(header_filename, output_filename_sorted)
if summary:
with open(summary_filename, 'a') as f:
f.write('Joint BAM file reheaded.\n')
# FIXME: check whether temp files are all deleted
if VERBOSE >= 1:
print 'Remove temporary files: adaID ' + adaID + ', fragment ' + frag_gen
remove_mapped_tempfiles(data_folder,
adaID,
frag_gen,
VERBOSE=VERBOSE,
rescue=rescue)
if summary:
with open(summary_filename, 'a') as f:
f.write('Temp mapping files removed.\n')
f.write('\n')
def premap_stampy(data_folder,
adaID,
VERBOSE=0,
threads=1,
summary=True,
maxreads=-1,
subsrate=0.05,
gapopen=40,
gapextend=3):
'''Call stampy for actual mapping'''
if VERBOSE:
print 'Premapping: adaID ', adaID
if summary:
summary_filename = get_premap_summary_filename(data_folder, adaID)
# Stampy can handle both gzipped and uncompressed fastq inputs
input_filenames = get_read_filenames(data_folder, adaID, gzip=True)
if not os.path.isfile(input_filenames[0]):
input_filenames = get_read_filenames(data_folder, adaID, gzip=False)
if not all(map(os.path.isfile, input_filenames)):
raise OSError('Input files for mapping not found: ' +
input_filenames[0])
# parallelize if requested
if threads == 1:
call_list = [
stampy_bin,
'--overwrite',
'-g',
get_reference_premap_index_filename(data_folder, adaID, ext=False),
'-h',
get_reference_premap_hash_filename(data_folder, adaID, ext=False),
'-o',
get_premapped_filename(data_folder, adaID, type='sam'),
'--insertsize=450',
'--insertsd=100',
'--substitutionrate=' + str(subsrate),
'--gapopen=' + str(gapopen),
'--gapextend=' + str(gapextend),
]
if maxreads > 0:
call_list.append('--numrecords=' + str(maxreads))
call_list.extend(['-M'] + input_filenames)
call_list = map(str, call_list)
if VERBOSE >= 2:
print ' '.join(call_list)
sp.call(call_list)
if summary:
with open(get_premap_summary_filename(data_folder, adaID),
'a') as f:
f.write('\nStampy premapped (single thread).\n')
# Convert to compressed BAM
convert_sam_to_bam(
get_premapped_filename(data_folder, adaID, type='bam'))
if summary:
with open(summary_filename, 'a') as f:
f.write('\nSAM file converted to compressed BAM: '+\
get_premapped_filename(data_folder, adaID, type='bam')+'\n')
else:
# Multithreading works as follows: call qsub + stampy, monitor the process
# IDs with qstat at regular intervals, and finally merge results with pysam
output_file_parts = [
get_premapped_filename(
data_folder, adaID, type='bam', part=(j + 1))
for j in xrange(threads)
]
# Submit map script
jobs_done = np.zeros(threads, bool)
job_IDs = np.zeros(threads, 'S30')
# Submit map call
import hivwholeseq
JOBDIR = hivwholeseq.__path__[0].rstrip('/') + '/'
JOBLOGOUT = JOBDIR + 'logout'
JOBLOGERR = JOBDIR + 'logerr'
cluster_time = ['23:59:59', '1:59:59']
vmem = '8G'
for j in xrange(threads):
call_list = [
'qsub', '-cwd', '-b', 'y', '-S', '/bin/bash', '-o', JOBLOGOUT,
'-e', JOBLOGERR, '-N', adaID + ' p' + str(j + 1), '-l',
'h_rt=' + cluster_time[threads >= 30], '-l', 'h_vmem=' + vmem,
stampy_bin, '--overwrite', '-g',
get_reference_premap_index_filename(
data_folder, adaID, ext=False), '-h',
get_reference_premap_hash_filename(
data_folder, adaID, ext=False), '-o',
get_premapped_filename(
data_folder, adaID, type='sam', part=(j + 1)),
'--processpart=' + str(j + 1) + '/' + str(threads),
'--insertsize=450', '--insertsd=100', '--substitutionrate=' +
str(subsrate), '--gapopen=' + str(gapopen),
'--gapextend=' + str(gapextend), '-M'
] + input_filenames
call_list = map(str, call_list)
if VERBOSE >= 2:
print ' '.join(call_list)
job_ID = sp.check_output(call_list)
job_ID = job_ID.split()[2]
job_IDs[j] = job_ID
# Monitor output
time_wait = 10 # secs
while not jobs_done.all():
# Sleep some time
time.sleep(time_wait)
# Get the output of qstat to check the status of jobs
qstat_output = sp.check_output(['qstat'])
qstat_output = qstat_output.split(
'\n')[:-1] # The last is an empty line
if VERBOSE >= 3:
print qstat_output
if len(qstat_output) < 3:
jobs_done[:] = True
break
else:
qstat_output = [line.split()[0] for line in qstat_output[2:]]
time_wait = 10 # secs
for j in xrange(threads):
if jobs_done[j]:
continue
if job_IDs[j] not in qstat_output:
# Convert to BAM for merging
if VERBOSE >= 1:
print 'Convert premapped reads to BAM for merging: adaID '+\
adaID+', part '+str(j+1)+ ' of '+ \
str(threads)
convert_sam_to_bam(output_file_parts[j])
# We do not need to wait if we did the conversion (it takes
# longer than some secs)
time_wait = 0
jobs_done[j] = True
if summary:
with open(summary_filename, 'a') as f:
f.write('Stampy premapped (' + str(threads) + ' threads).\n')
# Concatenate output files
if VERBOSE >= 1:
print 'Concatenate premapped reads: adaID ' + adaID + '...',
output_filename = get_premapped_filename(
data_folder, adaID, type='bam', unsorted=True)
pysam.cat('-o', output_filename, *output_file_parts)
if VERBOSE >= 1:
print 'done.'
if summary:
with open(summary_filename, 'a') as f:
f.write('BAM files concatenated (unsorted).\n')
# Sort the file by read names (to ensure the pair_generator)
# NOTE: we exclude the extension and the option -f because of a bug in samtools
if VERBOSE >= 1:
print 'Sort premapped reads: adaID ' + adaID
output_filename_sorted = get_premapped_filename(
data_folder, adaID, type='bam', unsorted=False)
pysam.sort('-n', output_filename, output_filename_sorted[:-4])
if summary:
with open(summary_filename, 'a') as f:
f.write('Joint BAM file sorted.\n')
# Reheader the file without BAM -> SAM -> BAM
if VERBOSE >= 1:
print 'Reheader premapped reads: adaID ' + adaID
header_filename = get_premapped_filename(
data_folder, adaID, type='sam', part=1)
pysam.reheader(header_filename, output_filename_sorted)
if summary:
with open(summary_filename, 'a') as f:
f.write('Joint BAM file reheaded.\n')
if VERBOSE >= 1:
print 'Remove temporary files: adaID ' + adaID
remove_premapped_tempfiles(data_folder, adaID, VERBOSE=VERBOSE)
if summary:
with open(summary_filename, 'a') as f:
f.write('Temp premapping files removed.\n')
f.write('\n')
def map_stampy_multithread(sample, fragment, VERBOSE=0, threads=2, summary=True,
filtered=True):
'''Map using stampy, multithread (via cluster requests, queueing race conditions possible)'''
import hivwholeseq
JOBDIR = hivwholeseq.__path__[0].rstrip('/')+'/'
JOBLOGOUT = JOBDIR+'logout/'
JOBLOGERR = JOBDIR+'logerr/'
cluster_time = ['23:59:59', '0:59:59']
vmem = '8G'
pname = patient.id
sample = patient.sample_table.loc[samplename]
seq_run = sample['run']
data_folder = MiSeq_runs[seq_run]['folder']
adaID = sample['adaID']
if VERBOSE:
print 'Map via stampy: '+pname+' '+samplename+' '+fragment
if summary:
summary_filename = get_map_initial_summary_filename(pname, samplename, fragment)
# Specific fragment (e.g. F5 --> F5bi)
frag_spec = filter(lambda x: fragment in x, sample['fragments'])
if not len(frag_spec):
raise ValueError(str(patient)+', '+samplename+': fragment '+fragment+' not found.')
frag_spec = frag_spec[0]
input_filename = get_input_filename(data_folder, adaID, frag_spec, type='bam')
# Submit map scripts in parallel to the cluster
jobs_done = np.zeros(threads, bool)
job_IDs = np.zeros(threads, 'S30')
for j in xrange(threads):
output_filename = get_mapped_to_initial_filename(pname, samplename,
fragment,
type='sam', part=(j+1))
# Map
call_list = ['qsub','-cwd',
'-b', 'y',
'-S', '/bin/bash',
'-o', JOBLOGOUT,
'-e', JOBLOGERR,
'-N', 'm '+samplename+fragment+' p'+str(j+1),
'-l', 'h_rt='+cluster_time[threads >= 10],
'-l', 'h_vmem='+vmem,
stampy_bin,
'--overwrite',
'-g', get_initial_index_filename(pname, fragment, ext=False),
'-h', get_initial_hash_filename(pname, fragment, ext=False),
'-o', output_filename,
'--processpart='+str(j+1)+'/'+str(threads),
'--substitutionrate='+subsrate,
'--gapopen', stampy_gapopen,
'--gapextend', stampy_gapextend]
if stampy_sensitive:
call_list.append('--sensitive')
call_list = call_list + ['-M', input_filename]
call_list = map(str, call_list)
if VERBOSE >= 2:
print ' '.join(call_list)
job_ID = sp.check_output(call_list)
job_ID = job_ID.split()[2]
job_IDs[j] = job_ID
# Monitor output
output_file_parts = [get_mapped_to_initial_filename(pname, samplename,
fragment,
type='bam', part=(j+1))
for j in xrange(threads)]
time_wait = 10 # secs
while not jobs_done.all():
# Sleep some time
time.sleep(time_wait)
# Get the output of qstat to check the status of jobs
qstat_output = sp.check_output(['qstat'])
qstat_output = qstat_output.split('\n')[:-1] # The last is an empty line
if len(qstat_output) < 3:
jobs_done[:] = True
break
else:
qstat_output = [line.split()[0] for line in qstat_output[2:]]
time_wait = 10 # secs
for j in xrange(threads):
if jobs_done[j]:
continue
if job_IDs[j] not in qstat_output:
# Convert to BAM for merging
if VERBOSE >= 1:
print 'Convert mapped reads to BAM for merging: sample '+\
samplename+', part '+str(j+1)+ ' of '+ \
str(threads)
convert_sam_to_bam(output_file_parts[j])
# We do not need to wait if we did the conversion (it takes
# longer than some secs)
time_wait = 0
jobs_done[j] = True
if summary:
with open(summary_filename, 'a') as f:
f.write('Stampy mapped ('+str(threads)+' threads).\n')
# Concatenate output files
output_filename = get_mapped_to_initial_filename(pname, samplename,
fragment,
type='bam', unsorted=True)
if VERBOSE >= 1:
print 'Concatenate premapped reads: sample '+samplename
pysam.cat('-o', output_filename, *output_file_parts)
if summary:
with open(summary_filename, 'a') as f:
f.write('BAM files concatenated (unsorted).\n')
# Sort the file by read names (to ensure the pair_generator)
output_filename_sorted = get_mapped_to_initial_filename(pname, samplename,
fragment,
type='bam')
# NOTE: we exclude the extension and the option -f because of a bug in samtools
if VERBOSE >= 1:
print 'Sort mapped reads: sample '+samplename
pysam.sort('-n', output_filename, output_filename_sorted[:-4])
if summary:
with open(summary_filename, 'a') as f:
f.write('Joint BAM file sorted.\n')
# Reheader the file without BAM -> SAM -> BAM
if VERBOSE >= 1:
print 'Reheader mapped reads: sample '+samplename
header_filename = get_mapped_to_initial_filename(pname, samplename,
fragment,
type='sam', part=1)
pysam.reheader(header_filename, output_filename_sorted)
if summary:
with open(summary_filename, 'a') as f:
f.write('Joint BAM file reheaded.\n')
if VERBOSE >= 1:
print 'Remove temporary files: sample '+samplename
remove_mapped_init_tempfiles(pname, samplename, fragment, VERBOSE=VERBOSE)
if summary:
with open(summary_filename, 'a') as f:
f.write('Temp mapping files removed.\n')
f.write('\n')