def __init__(self, ref_prefix, max_n):
self.max_n = max_n # use up to max_n gram
# reference files
self.files = self.get_ref_files(ref_prefix)
# number of references per file
self.nref = count_lines(self.files[0])
for filename in self.files:
n = count_lines(filename)
assert self.nref == n, '%s has %s lines' % (filename, n)
# counters for ngrams
self.counters = [RefCounter(max_n) for i in range(self.nref)]
self.load()
def __init__(self,
ffilename,
efilename,
afilename,
outputdir,
alpha,
threshold,
length_factor=False,
lexical_weighter=None,
maximize_derivation=False):
self.ffilename = ffilename
self.efilename = efilename
self.afilename = afilename
self.outputdir = outputdir
self.alpha = alpha
self.threshold = threshold
self.length_factor = length_factor
self.lexical_weighter = lexical_weighter
self.maximize_derivation = maximize_derivation
self.counter = RuleCounter()
self.corpus_size = count_lines(ffilename)
system('rm -rf %s' % outputdir)
system('mkdir %s' % outputdir)
def read_table(json_table_path):
with open(json_table_path) as table_file:
tables = {}
for line in tqdm(table_file, total=count_lines(json_table_path)):
d = json.loads(line)
tables[d['id']] = d
return tables
def __init__(self, total=-1, input='', file=stdout):
assert total == -1 or input == '', \
"user should specify either 'total' or 'input'"
if total != -1:
self.total = total
elif input:
self.total = count_lines(input)
else:
assert False, "please specify either 'total' or 'input'"
self.percent = 0
self.file = file
def upload(self, uploader):
# Information about called peaks
n_spp_peaks = common.count_lines(self.peaks_fn)
print "%s peaks called by spp" % n_spp_peaks
print "%s of those peaks removed due to bad coordinates" % (n_spp_peaks - common.count_lines(self.fixed_peaks_fn))
print "First 50 peaks"
print subprocess.check_output('head -50 %s' % self.fixed_peaks_fn, shell=True, stderr=subprocess.STDOUT)
# Upload bigBed if applicable
if self.bigbed:
self.peaks_bb_fn = common.bed2bb(self.fixed_peaks_fn, self.chrom_sizes.name, self.as_file.name)
if self.peaks_bb_fn:
self.peaks_bb = uploader.upload(self.peaks_bb_fn)
if not filecmp.cmp(self.peaks_fn, self.fixed_peaks_fn): print "Returning peaks with fixed coordinates"
# Upload peaks
print subprocess.check_output(shlex.split("gzip %s" % self.fixed_peaks_fn))
self.peaks = uploader.upload(self.fixed_peaks_fn + ".gz")
# Upload cross-correlations
self.xcor_plot = uploader.upload(self.xcor_plot)
self.xcor_scores = uploader.upload(self.xcor_scores)
def rescale_scores(fn, scores_col, new_min=10, new_max=1000):
n_peaks = common.count_lines(fn)
sorted_fn = 'sorted-%s' %(fn)
rescaled_fn = 'rescaled-%s' %(fn)
out,err = common.run_pipe([
'sort -k %dgr,%dgr %s' %(scores_col, scores_col, fn),
r"""awk 'BEGIN{FS="\t";OFS="\t"}{if (NF != 0) print $0}'"""],
sorted_fn)
out, err = common.run_pipe([
'head -n 1 %s' %(sorted_fn),
'cut -f %s' %(scores_col)])
max_score = float(out.strip())
out, err = common.run_pipe([
'tail -n 1 %s' %(sorted_fn),
'cut -f %s' %(scores_col)])
min_score = float(out.strip())
out,err = common.run_pipe([
'cat %s' %(sorted_fn),
r"""awk 'BEGIN{OFS="\t"}{n=$%d;a=%d;b=%d;x=%d;y=%d}""" %(scores_col, min_score, max_score, new_min, new_max) + \
r"""{$%d=int(((n-a)*(y-x)/(b-a))+x) ; print $0}'""" %(scores_col)],
rescaled_fn)
return rescaled_fn
def rescale_scores(fn, scores_col, new_min=10, new_max=1000):
n_peaks = common.count_lines(fn)
sorted_fn = 'sorted-%s' % (fn)
rescaled_fn = 'rescaled-%s' % (fn)
out, err = common.run_pipe([
'sort -k %dgr,%dgr %s' % (scores_col, scores_col, fn),
r"""awk 'BEGIN{FS="\t";OFS="\t"}{if (NF != 0) print $0}'"""
], sorted_fn)
out, err = common.run_pipe(
['head -n 1 %s' % (sorted_fn),
'cut -f %s' % (scores_col)])
max_score = float(out.strip())
out, err = common.run_pipe(
['tail -n 1 %s' % (sorted_fn),
'cut -f %s' % (scores_col)])
min_score = float(out.strip())
out,err = common.run_pipe([
'cat %s' %(sorted_fn),
r"""awk 'BEGIN{OFS="\t"}{n=$%d;a=%d;b=%d;x=%d;y=%d}""" %(scores_col, min_score, max_score, new_min, new_max) + \
r"""{$%d=int(((n-a)*(y-x)/(b-a))+x) ; print $0}'""" %(scores_col)],
rescaled_fn)
return rescaled_fn
def internal_pseudoreplicate_overlap(rep1_peaks,
rep2_peaks,
pooled_peaks,
rep1_ta,
rep1_xcor,
paired_end,
chrom_sizes,
as_file,
peak_type,
prefix,
fragment_length=None):
rep1_peaks_file = dxpy.DXFile(rep1_peaks)
rep2_peaks_file = dxpy.DXFile(rep2_peaks)
pooled_peaks_file = dxpy.DXFile(pooled_peaks)
rep1_ta_file = dxpy.DXFile(rep1_ta)
rep1_xcor_file = dxpy.DXFile(rep1_xcor)
chrom_sizes_file = dxpy.DXFile(chrom_sizes)
as_file_file = dxpy.DXFile(as_file)
# Input filenames - necessary to define each explicitly because input files
# could have the same name, in which case subsequent
# file would overwrite previous file
rep1_peaks_fn = 'rep1-%s' % (rep1_peaks_file.name)
rep2_peaks_fn = 'rep2-%s' % (rep2_peaks_file.name)
pooled_peaks_fn = 'pooled-%s' % (pooled_peaks_file.name)
rep1_ta_fn = 'r1ta_%s' % (rep1_ta_file.name)
rep1_xcor_fn = 'r1xc_%s' % (rep1_xcor_file.name)
chrom_sizes_fn = 'chrom.sizes'
as_file_fn = '%s.as' % (peak_type)
# Output filenames
if prefix:
basename = prefix
else:
# strip off the peak and compression extensions
m = re.match('(.*)(\.%s)+(\.((gz)|(Z)|(bz)|(bz2)))' % (peak_type),
pooled_peaks.name)
if m:
basename = m.group(1)
else:
basename = pooled_peaks.name
overlapping_peaks_fn = '%s.replicated.%s' % (basename, peak_type)
overlapping_peaks_bb_fn = overlapping_peaks_fn + '.bb'
rejected_peaks_fn = '%s.rejected.%s' % (basename, peak_type)
rejected_peaks_bb_fn = rejected_peaks_fn + '.bb'
# Intermediate filenames
overlap_tr_fn = 'replicated_tr.%s' % (peak_type)
overlap_pr_fn = 'replicated_pr.%s' % (peak_type)
# Download file inputs to the local file system with local filenames
dxpy.download_dxfile(rep1_peaks_file.get_id(), rep1_peaks_fn)
dxpy.download_dxfile(rep2_peaks_file.get_id(), rep2_peaks_fn)
dxpy.download_dxfile(pooled_peaks_file.get_id(), pooled_peaks_fn)
dxpy.download_dxfile(rep1_ta_file.get_id(), rep1_ta_fn)
dxpy.download_dxfile(rep1_xcor_file.get_id(), rep1_xcor_fn)
dxpy.download_dxfile(chrom_sizes_file.get_id(), chrom_sizes_fn)
dxpy.download_dxfile(as_file_file.get_id(), as_file_fn)
logger.info(subprocess.check_output('set -x; ls -l', shell=True))
# the only difference between the peak_types is how the extra columns are
# handled
if peak_type == "narrowPeak":
awk_command = r"""awk 'BEGIN{FS="\t";OFS="\t"}{s1=$3-$2; s2=$13-$12; if (($21/s1 >= 0.5) || ($21/s2 >= 0.5)) {print $0}}'"""
cut_command = 'cut -f 1-10'
bed_type = 'bed6+4'
elif peak_type == "gappedPeak":
awk_command = r"""awk 'BEGIN{FS="\t";OFS="\t"}{s1=$3-$2; s2=$18-$17; if (($31/s1 >= 0.5) || ($31/s2 >= 0.5)) {print $0}}'"""
cut_command = 'cut -f 1-15'
bed_type = 'bed12+3'
elif peak_type == "broadPeak":
awk_command = r"""awk 'BEGIN{FS="\t";OFS="\t"}{s1=$3-$2; s2=$12-$11; if (($19/s1 >= 0.5) || ($19/s2 >= 0.5)) {print $0}}'"""
cut_command = 'cut -f 1-9'
bed_type = 'bed6+3'
else:
assert peak_type in [
'narrowPeak', 'gappedPeak', 'broadPeak'
], "%s is unrecognized. peak_type should be narrowPeak, gappedPeak or broadPeak." % (
peak_type)
# Find pooled peaks that overlap Rep1 and Rep2 where overlap is defined as
# the fractional overlap wrt any one of the overlapping peak pairs > 0.5
out, err = common.run_pipe([
'intersectBed -wo -a %s -b %s' %
(pooled_peaks_fn, rep1_peaks_fn), awk_command, cut_command, 'sort -u',
'intersectBed -wo -a stdin -b %s' %
(rep2_peaks_fn), awk_command, cut_command, 'sort -u'
], overlap_tr_fn)
print("%d peaks overlap with both true replicates" %
(common.count_lines(overlap_tr_fn)))
# this is a simplicate analysis
# overlapping peaks are just based on pseudoreps of the one pool
out, err = common.run_pipe(['cat %s' % (overlap_tr_fn), 'sort -u'],
overlapping_peaks_fn)
print("%d peaks overlap" % (common.count_lines(overlapping_peaks_fn)))
# rejected peaks
out, err = common.run_pipe([
'intersectBed -wa -v -a %s -b %s' %
(pooled_peaks_fn, overlapping_peaks_fn)
], rejected_peaks_fn)
print("%d peaks were rejected" % (common.count_lines(rejected_peaks_fn)))
# calculate FRiP (Fraction of Reads in Peaks)
# Extract the fragment length estimate from column 3 of the
# cross-correlation scores file or use the user-defined
# fragment_length if given.
if fragment_length is not None:
fraglen = fragment_length
fragment_length_given_by_user = True
else:
fraglen = common.xcor_fraglen(rep1_xcor_fn)
fragment_length_given_by_user = False
# FRiP
reads_in_peaks_fn = 'reads_in_%s.ta' % (peak_type)
n_reads, n_reads_in_peaks, frip_score = common.frip(
rep1_ta_fn,
rep1_xcor_fn,
overlapping_peaks_fn,
chrom_sizes_fn,
fraglen,
reads_in_peaks_fn=reads_in_peaks_fn)
# count peaks
npeaks_in = common.count_lines(common.uncompress(pooled_peaks_fn))
npeaks_out = common.count_lines(overlapping_peaks_fn)
npeaks_rejected = common.count_lines(rejected_peaks_fn)
# make bigBed files for visualization
overlapping_peaks_bb_fn = common.bed2bb(overlapping_peaks_fn,
chrom_sizes_fn,
as_file_fn,
bed_type=bed_type)
rejected_peaks_bb_fn = common.bed2bb(rejected_peaks_fn,
chrom_sizes_fn,
as_file_fn,
bed_type=bed_type)
# Upload file outputs from the local file system.
overlapping_peaks = dxpy.upload_local_file(
common.compress(overlapping_peaks_fn))
overlapping_peaks_bb = dxpy.upload_local_file(overlapping_peaks_bb_fn)
rejected_peaks = dxpy.upload_local_file(common.compress(rejected_peaks_fn))
rejected_peaks_bb = dxpy.upload_local_file(rejected_peaks_bb_fn)
output = {
"overlapping_peaks": dxpy.dxlink(overlapping_peaks),
"overlapping_peaks_bb": dxpy.dxlink(overlapping_peaks_bb),
"rejected_peaks": dxpy.dxlink(rejected_peaks),
"rejected_peaks_bb": dxpy.dxlink(rejected_peaks_bb),
"npeaks_in": npeaks_in,
"npeaks_out": npeaks_out,
"npeaks_rejected": npeaks_rejected,
"frip_nreads": n_reads,
"frip_nreads_in_peaks": n_reads_in_peaks,
"frip_score": frip_score,
"fragment_length_used": fraglen,
"fragment_length_given_by_user": fragment_length_given_by_user
}
return output
def replicated_overlap(rep1_peaks,
rep2_peaks,
pooled_peaks,
pooledpr1_peaks,
pooledpr2_peaks,
rep1_ta,
rep1_xcor,
rep2_ta,
rep2_xcor,
paired_end,
chrom_sizes,
as_file,
peak_type,
prefix,
fragment_length=None):
rep1_peaks_file = dxpy.DXFile(rep1_peaks)
rep2_peaks_file = dxpy.DXFile(rep2_peaks)
pooled_peaks_file = dxpy.DXFile(pooled_peaks)
pooledpr1_peaks_file = dxpy.DXFile(pooledpr1_peaks)
pooledpr2_peaks_file = dxpy.DXFile(pooledpr2_peaks)
rep1_ta_file = dxpy.DXFile(rep1_ta)
rep2_ta_file = dxpy.DXFile(rep2_ta)
rep1_xcor_file = dxpy.DXFile(rep1_xcor)
rep2_xcor_file = dxpy.DXFile(rep2_xcor)
chrom_sizes_file = dxpy.DXFile(chrom_sizes)
as_file_file = dxpy.DXFile(as_file)
# Input filenames - necessary to define each explicitly because input files
# could have the same name, in which case subsequent
# file would overwrite previous file
rep1_peaks_fn = 'rep1-%s' % (rep1_peaks_file.name)
rep2_peaks_fn = 'rep2-%s' % (rep2_peaks_file.name)
pooled_peaks_fn = 'pooled-%s' % (pooled_peaks_file.name)
pooledpr1_peaks_fn = 'pooledpr1-%s' % (pooledpr1_peaks_file.name)
pooledpr2_peaks_fn = 'pooledpr2-%s' % (pooledpr2_peaks_file.name)
rep1_ta_fn = 'r1ta_%s' % (rep1_ta_file.name)
rep2_ta_fn = 'r2ta_%s' % (rep2_ta_file.name)
rep1_xcor_fn = 'r1cc_%s' % (rep1_xcor_file.name)
rep2_xcor_fn = 'r2cc_%s' % (rep2_xcor_file.name)
chrom_sizes_fn = 'chrom.sizes'
as_file_fn = '%s.as' % (peak_type)
# Output filenames
if prefix:
basename = prefix
else:
# strip off the peak and compression extensions
m = re.match('(.*)(\.%s)+(\.((gz)|(Z)|(bz)|(bz2)))' % (peak_type),
pooled_peaks.name)
if m:
basename = m.group(1)
else:
basename = pooled_peaks.name
overlapping_peaks_fn = '%s.replicated.%s' % (basename, peak_type)
overlapping_peaks_bb_fn = overlapping_peaks_fn + '.bb'
rejected_peaks_fn = '%s.rejected.%s' % (basename, peak_type)
rejected_peaks_bb_fn = rejected_peaks_fn + '.bb'
# Intermediate filenames
overlap_tr_fn = 'replicated_tr.%s' % (peak_type)
overlap_pr_fn = 'replicated_pr.%s' % (peak_type)
# Download file inputs to the local file system with local filenames
dxpy.download_dxfile(rep1_peaks_file.get_id(), rep1_peaks_fn)
dxpy.download_dxfile(rep2_peaks_file.get_id(), rep2_peaks_fn)
dxpy.download_dxfile(pooled_peaks_file.get_id(), pooled_peaks_fn)
dxpy.download_dxfile(pooledpr1_peaks_file.get_id(), pooledpr1_peaks_fn)
dxpy.download_dxfile(pooledpr2_peaks_file.get_id(), pooledpr2_peaks_fn)
dxpy.download_dxfile(rep1_ta_file.get_id(), rep1_ta_fn)
dxpy.download_dxfile(rep2_ta_file.get_id(), rep2_ta_fn)
dxpy.download_dxfile(rep1_xcor_file.get_id(), rep1_xcor_fn)
dxpy.download_dxfile(rep2_xcor_file.get_id(), rep2_xcor_fn)
dxpy.download_dxfile(chrom_sizes_file.get_id(), chrom_sizes_fn)
dxpy.download_dxfile(as_file_file.get_id(), as_file_fn)
pool_applet = dxpy.find_one_data_object(classname='applet',
name='pool',
project=dxpy.PROJECT_CONTEXT_ID,
zero_ok=False,
more_ok=False,
return_handler=True)
pool_replicates_subjob = \
pool_applet.run(
{"inputs": [rep1_ta, rep2_ta],
"prefix": 'pooled_reps'},
name='Pool replicates')
# If fragment length was given by user we skip pooled_replicates
# _xcor_subjob, set the pool_xcor_filename to None, and update
# the flag fragment_length_given_by_user. Otherwise, run the subjob
# to be able to extract the fragment length fron cross-correlations.
if fragment_length is not None:
pool_xcor_filename = None
fraglen = fragment_length
fragment_length_given_by_user = True
else:
pooled_replicates_xcor_subjob = \
xcor_only(
pool_replicates_subjob.get_output_ref("pooled"),
paired_end,
spp_version=None,
name='Pool cross-correlation')
pooled_replicates_xcor_subjob.wait_on_done()
pool_xcor_link = pooled_replicates_xcor_subjob.describe(
)['output'].get("CC_scores_file")
pool_xcor_file = dxpy.get_handler(pool_xcor_link)
pool_xcor_filename = 'poolcc_%s' % (pool_xcor_file.name)
dxpy.download_dxfile(pool_xcor_file.get_id(), pool_xcor_filename)
fraglen = common.xcor_fraglen(pool_xcor_filename)
fragment_length_given_by_user = False
pool_replicates_subjob.wait_on_done()
pool_ta_link = pool_replicates_subjob.describe()['output'].get("pooled")
pool_ta_file = dxpy.get_handler(pool_ta_link)
pool_ta_filename = 'poolta_%s' % (pool_ta_file.name)
dxpy.download_dxfile(pool_ta_file.get_id(), pool_ta_filename)
logger.info(subprocess.check_output('set -x; ls -l', shell=True))
# the only difference between the peak_types is how the extra columns are
# handled
if peak_type == "narrowPeak":
awk_command = r"""awk 'BEGIN{FS="\t";OFS="\t"}{s1=$3-$2; s2=$13-$12; if (($21/s1 >= 0.5) || ($21/s2 >= 0.5)) {print $0}}'"""
cut_command = 'cut -f 1-10'
bed_type = 'bed6+4'
elif peak_type == "gappedPeak":
awk_command = r"""awk 'BEGIN{FS="\t";OFS="\t"}{s1=$3-$2; s2=$18-$17; if (($31/s1 >= 0.5) || ($31/s2 >= 0.5)) {print $0}}'"""
cut_command = 'cut -f 1-15'
bed_type = 'bed12+3'
elif peak_type == "broadPeak":
awk_command = r"""awk 'BEGIN{FS="\t";OFS="\t"}{s1=$3-$2; s2=$12-$11; if (($19/s1 >= 0.5) || ($19/s2 >= 0.5)) {print $0}}'"""
cut_command = 'cut -f 1-9'
bed_type = 'bed6+3'
else:
assert peak_type in [
'narrowPeak', 'gappedPeak', 'broadPeak'
], "%s is unrecognized. peak_type should be narrowPeak, gappedPeak or broadPeak." % (
peak_type)
# Find pooled peaks that overlap Rep1 and Rep2 where overlap is defined as
# the fractional overlap wrt any one of the overlapping peak pairs > 0.5
out, err = common.run_pipe([
'intersectBed -wo -a %s -b %s' %
(pooled_peaks_fn, rep1_peaks_fn), awk_command, cut_command, 'sort -u',
'intersectBed -wo -a stdin -b %s' %
(rep2_peaks_fn), awk_command, cut_command, 'sort -u'
], overlap_tr_fn)
print("%d peaks overlap with both true replicates" %
(common.count_lines(overlap_tr_fn)))
# Find pooled peaks that overlap PseudoRep1 and PseudoRep2 where
# overlap is defined as the fractional overlap wrt any one of the
# overlapping peak pairs > 0.5
out, err = common.run_pipe([
'intersectBed -wo -a %s -b %s' % (pooled_peaks_fn, pooledpr1_peaks_fn),
awk_command, cut_command, 'sort -u',
'intersectBed -wo -a stdin -b %s' %
(pooledpr2_peaks_fn), awk_command, cut_command, 'sort -u'
], overlap_pr_fn)
print("%d peaks overlap with both pooled pseudoreplicates" %
(common.count_lines(overlap_pr_fn)))
# Combine peak lists
out, err = common.run_pipe(
['cat %s %s' % (overlap_tr_fn, overlap_pr_fn), 'sort -u'],
overlapping_peaks_fn)
print(
"%d peaks overlap with true replicates or with pooled pseudoreplicates"
% (common.count_lines(overlapping_peaks_fn)))
# rejected peaks
out, err = common.run_pipe([
'intersectBed -wa -v -a %s -b %s' %
(pooled_peaks_fn, overlapping_peaks_fn)
], rejected_peaks_fn)
print("%d peaks were rejected" % (common.count_lines(rejected_peaks_fn)))
# calculate FRiP (Fraction of Reads in Peaks)
reads_in_peaks_fn = 'reads_in_%s.ta' % (peak_type)
n_reads, n_reads_in_peaks, frip_score = common.frip(
pool_ta_filename,
pool_xcor_filename,
overlapping_peaks_fn,
chrom_sizes_fn,
fraglen,
reads_in_peaks_fn=reads_in_peaks_fn)
# count peaks
npeaks_in = common.count_lines(common.uncompress(pooled_peaks_fn))
npeaks_out = common.count_lines(overlapping_peaks_fn)
npeaks_rejected = common.count_lines(rejected_peaks_fn)
# make bigBed files for visualization
overlapping_peaks_bb_fn = common.bed2bb(overlapping_peaks_fn,
chrom_sizes_fn,
as_file_fn,
bed_type=bed_type)
rejected_peaks_bb_fn = common.bed2bb(rejected_peaks_fn,
chrom_sizes_fn,
as_file_fn,
bed_type=bed_type)
# Upload file outputs from the local file system.
overlapping_peaks = dxpy.upload_local_file(
common.compress(overlapping_peaks_fn))
overlapping_peaks_bb = dxpy.upload_local_file(overlapping_peaks_bb_fn)
rejected_peaks = dxpy.upload_local_file(common.compress(rejected_peaks_fn))
rejected_peaks_bb = dxpy.upload_local_file(rejected_peaks_bb_fn)
output = {
"overlapping_peaks": dxpy.dxlink(overlapping_peaks),
"overlapping_peaks_bb": dxpy.dxlink(overlapping_peaks_bb),
"rejected_peaks": dxpy.dxlink(rejected_peaks),
"rejected_peaks_bb": dxpy.dxlink(rejected_peaks_bb),
"npeaks_in": npeaks_in,
"npeaks_out": npeaks_out,
"npeaks_rejected": npeaks_rejected,
"frip_nreads": n_reads,
"frip_nreads_in_peaks": n_reads_in_peaks,
"frip_score": frip_score,
"fragment_length_used": fraglen,
"fragment_length_given_by_user": fragment_length_given_by_user
}
return output
def main(experiment,
control,
xcor_scores_input,
npeaks,
nodups,
bigbed,
chrom_sizes,
spp_version,
as_file=None,
prefix=None,
fragment_length=None):
# The following line(s) initialize your data object inputs on the platform
# into dxpy.DXDataObject instances that you can start using immediately.
experiment_file = dxpy.DXFile(experiment)
control_file = dxpy.DXFile(control)
xcor_scores_input_file = dxpy.DXFile(xcor_scores_input)
chrom_sizes_file = dxpy.DXFile(chrom_sizes)
chrom_sizes_filename = chrom_sizes_file.name
dxpy.download_dxfile(chrom_sizes_file.get_id(), chrom_sizes_filename)
if bigbed:
as_file_file = dxpy.DXFile(as_file)
as_file_filename = as_file_file.name
dxpy.download_dxfile(as_file_file.get_id(), as_file_filename)
# The following line(s) download your file inputs to the local file system
# using variable names for the filenames.
experiment_filename = experiment_file.name
dxpy.download_dxfile(experiment_file.get_id(), experiment_filename)
control_filename = control_file.name
dxpy.download_dxfile(control_file.get_id(), control_filename)
xcor_scores_input_filename = xcor_scores_input_file.name
dxpy.download_dxfile(xcor_scores_input_file.get_id(),
xcor_scores_input_filename)
if not prefix:
output_filename_prefix = \
experiment_filename.rstrip('.gz').rstrip('.tagAlign')
else:
output_filename_prefix = prefix
peaks_filename = output_filename_prefix + '.regionPeak'
# spp adds .gz, so this is the file name that's actually created
final_peaks_filename = peaks_filename + '.gz'
xcor_plot_filename = output_filename_prefix + '.pdf'
xcor_scores_filename = output_filename_prefix + '.ccscores'
logger.info(
subprocess.check_output('ls -l', shell=True, stderr=subprocess.STDOUT))
# third column in the cross-correlation scores input file
# if fragment_length is provided, use that. Else read
# fragment length from xcor file
if fragment_length is not None:
fraglen = str(fragment_length)
logger.info("User given fragment length %s" % (fraglen))
else:
fraglen_column = 3
with open(xcor_scores_input_filename, 'r') as f:
line = f.readline()
fraglen = line.split('\t')[fraglen_column - 1]
logger.info("Read fragment length: %s" % (fraglen))
# spp_tarball = SPP_VERSION_MAP.get(spp_version)
# assert spp_tarball, "spp version %s is not supported" % (spp_version)
# install spp
# subprocess.check_output(shlex.split('R CMD INSTALL %s' % (spp_tarball)))
run_spp = '/phantompeakqualtools/run_spp.R'
spp_command = (
"Rscript %s -p=%d -c=%s -i=%s -npeak=%d -speak=%s -savr=%s -savp=%s -rf -out=%s"
% (run_spp, cpu_count(), experiment_filename, control_filename, npeaks,
fraglen, peaks_filename, xcor_plot_filename, xcor_scores_filename))
logger.info(spp_command)
subprocess.check_call(shlex.split(spp_command))
# when one of the peak coordinates are an exact multiple of 10, spp (R)
# outputs the coordinate in scientific notation
# this changes any such coodinates to decimal notation
# this assumes 10-column output and that the 2nd and 3rd columns are
# coordinates
# the ($2>0)?$2:0) is needed because spp sometimes calls peaks with a
# negative start coordinate (particularly chrM) and will cause slopBed
# to halt at that line, truncating the output of the pipe
# slopBed adjusts feature end coordinates that go off the end of the
# chromosome
# bedClip removes any features that are still not within the boundaries of
# the chromosome
fix_coordinate_peaks_filename = \
output_filename_prefix + '.fixcoord.regionPeak'
out, err = common.run_pipe([
"gzip -dc %s" % (final_peaks_filename),
"tee %s" % (peaks_filename),
r"""awk 'BEGIN{OFS="\t"}{print $1,sprintf("%i",($2>0)?$2:0),sprintf("%i",$3),$4,$5,$6,$7,$8,$9,$10}'""",
'slopBed -i stdin -g %s -b 0' % (chrom_sizes_filename),
'bedClip stdin %s %s' %
(chrom_sizes_filename, fix_coordinate_peaks_filename)
])
# These lines transfer the peaks files to the temporary workspace for
# debugging later
# Only at the end are the final files uploaded that will be returned from
# the applet
dxpy.upload_local_file(peaks_filename)
dxpy.upload_local_file(fix_coordinate_peaks_filename)
n_spp_peaks = common.count_lines(peaks_filename)
logger.info("%s peaks called by spp" % (n_spp_peaks))
logger.info(
"%s of those peaks removed due to bad coordinates" %
(n_spp_peaks - common.count_lines(fix_coordinate_peaks_filename)))
print("First 50 peaks")
subprocess.check_output('head -50 %s' % (fix_coordinate_peaks_filename),
shell=True)
if bigbed:
peaks_bb_filename = \
common.bed2bb(fix_coordinate_peaks_filename, chrom_sizes_filename, as_file_filename)
if peaks_bb_filename:
peaks_bb = dxpy.upload_local_file(peaks_bb_filename)
if not filecmp.cmp(peaks_filename, fix_coordinate_peaks_filename):
logger.info("Returning peaks with fixed coordinates")
subprocess.check_call(
shlex.split('gzip -n %s' % (fix_coordinate_peaks_filename)))
final_peaks_filename = fix_coordinate_peaks_filename + '.gz'
subprocess.check_call('ls -l', shell=True)
# print subprocess.check_output('head %s' %(final_peaks_filename), shell=True, stderr=subprocess.STDOUT)
# print subprocess.check_output('head %s' %(xcor_scores_filename), shell=True, stderr=subprocess.STDOUT)
peaks = dxpy.upload_local_file(final_peaks_filename)
xcor_plot = dxpy.upload_local_file(xcor_plot_filename)
xcor_scores = dxpy.upload_local_file(xcor_scores_filename)
output = {}
output["peaks"] = dxpy.dxlink(peaks)
output["xcor_plot"] = dxpy.dxlink(xcor_plot)
output["xcor_scores"] = dxpy.dxlink(xcor_scores)
if bigbed and peaks_bb_filename:
output["peaks_bb"] = dxpy.dxlink(peaks_bb)
return output
def read_and_write_query(query_question_path,
tables,
question_output_path,
sql_output_path,
table_path,
valid_file_counter,
debug=False,
do_append=False):
sql_parser = SQLParser()
if do_append:
sql_writer = open(sql_output_path, 'a')
question_writer = open(question_output_path, 'a')
table_writer = open(table_path, 'a')
else:
sql_writer = open(sql_output_path, 'w')
question_writer = open(question_output_path, 'w')
table_writer = open(table_path, 'w')
num_of_unicode_error = 0
num_of_non_parsable_error = 0
with open(query_question_path) as qq_file:
queries = []
questions = []
counter = 0
for line in tqdm(qq_file, total=count_lines(query_question_path)):
data = json.loads(line)
question = data['question']
table_id = data['table_id']
table = tables[table_id]
column_names = table["header"]
#print(column_names)
sql = data['sql']
select_col = table["header"][int(sql["sel"])]
agg = agg_ops[int(sql["agg"])]
conditions = sql["conds"]
use_column_name = True
query = Query(int(sql["sel"]), int(sql["agg"]), column_names,
use_column_name, conditions)
#print("select col: " + select_col)
#print("agg: " + agg)
#print(question)
#print(sql)
#print(col_names)
hasError = False
try:
sql_query = query.__repr__()
col_names = " COL_END COL_START ".join(
str(x) for x in column_names)
except:
if debug:
print("ERROR in line unicode" + str(counter))
hasError = True
num_of_unicode_error += 1
if not hasError:
try:
#new_query, orig_table_name = fix_table_name(query)
parse_tree, rule_list = sql_parser.parse(sql_query,
get_rules=True)
sql_writer.write(sql_query + "\n")
question_writer.write(question + " COL_START " +
col_names + " COL_END\n")
valid_file_counter += 1
except:
if debug:
print("ERROR in line " + str(counter) + " :" +
str(sql_query))
num_of_non_parsable_error += 1
counter += 1
#if counter == 10:
# break
print("Unicode error: " + str(num_of_unicode_error))
print("Nonparsable error: " + str(num_of_non_parsable_error))
return valid_file_counter
def main(rep1_ta,
ctl1_ta,
rep1_xcor,
rep1_paired_end,
npeaks,
nodups,
chrom_sizes,
spp_version,
rep2_ta=None,
ctl2_ta=None,
rep2_xcor=None,
rep2_paired_end=None,
as_file=None,
idr_peaks=False,
fragment_length=None,
spp_instance=None):
rep1_ta_file = dxpy.DXFile(rep1_ta)
dxpy.download_dxfile(rep1_ta_file.get_id(), rep1_ta_file.name)
rep1_ta_filename = rep1_ta_file.name
ntags_rep1 = common.count_lines(rep1_ta_filename)
simplicate_experiment = rep1_ta and not rep2_ta
if simplicate_experiment:
logger.info(
"No rep2 tags specified so processing as a simplicate experiment.")
else:
logger.info(
"Rep1 and rep2 tags specified so processing as a replicated experiment."
)
if not simplicate_experiment:
assert rep1_paired_end == rep2_paired_end, 'Mixed PE/SE not supported'
rep2_ta_file = dxpy.DXFile(rep2_ta)
dxpy.download_dxfile(rep2_ta_file.get_id(), rep2_ta_file.name)
rep2_ta_filename = rep2_ta_file.name
ntags_rep2 = common.count_lines(rep2_ta_filename)
paired_end = rep1_paired_end
unary_control = (ctl1_ta == ctl2_ta) or not ctl2_ta
ctl1_ta_file = dxpy.DXFile(ctl1_ta)
dxpy.download_dxfile(ctl1_ta_file.get_id(), ctl1_ta_file.name)
ctl1_ta_filename = ctl1_ta_file.name
if not unary_control:
ctl2_ta_file = dxpy.DXFile(ctl2_ta)
dxpy.download_dxfile(ctl2_ta_file.get_id(), ctl2_ta_file.name)
ctl2_ta_filename = ctl2_ta_file.name
else:
ctl2_ta_file = ctl1_ta_file
ctl2_ta_filename = ctl1_ta_file.name
ntags_ctl1 = common.count_lines(ctl1_ta_filename)
ntags_ctl2 = common.count_lines(ctl2_ta_filename)
rep1_control = ctl1_ta # default. May be changed later.
rep1_ctl_msg = "control rep1"
rep2_control = ctl2_ta # default. May be changed later.
rep2_ctl_msg = "control rep2"
rep_info = [(ntags_rep1, 'replicate 1', rep1_ta_filename)]
if not simplicate_experiment:
rep_info.append((ntags_rep2, 'replicate 2', rep2_ta_filename))
rep_info.extend([(ntags_ctl1, 'control 1', ctl1_ta_filename),
(ntags_ctl2, 'control 2', ctl2_ta_filename)])
for n, name, filename in rep_info:
logger.info("Found %d tags in %s file %s" % (n, name, filename))
subprocess.check_output('ls -l', shell=True, stderr=subprocess.STDOUT)
if not simplicate_experiment:
pool_applet = dxpy.find_one_data_object(
classname='applet',
name='pool',
project=dxpy.PROJECT_CONTEXT_ID,
zero_ok=False,
more_ok=False,
return_handler=True)
pool_replicates_subjob = \
pool_applet.run(
{"inputs": [rep1_ta, rep2_ta],
"prefix": 'pooled_reps'},
name='Pool replicates')
pooled_replicates = pool_replicates_subjob.get_output_ref("pooled")
pooled_replicates_xcor_subjob = \
xcor_only(
pooled_replicates,
paired_end,
spp_version,
name='Pool cross-correlation')
if unary_control:
logger.info("Only one control supplied.")
if not simplicate_experiment:
logger.info(
"Using one control for both replicate 1 and 2 and for the pool."
)
rep2_control = rep1_control
control_for_pool = rep1_control
pool_ctl_msg = "one control"
else:
pool_controls_subjob = pool_applet.run(
{
"inputs": [ctl1_ta, ctl2_ta],
"prefix": "PL_ctls"
},
name='Pool controls')
pooled_controls = pool_controls_subjob.get_output_ref("pooled")
# always use the pooled controls for the pool
control_for_pool = pooled_controls
pool_ctl_msg = "pooled controls"
# use the pooled controls for the reps depending on the ratio of rep to
# control reads
ratio_ctl_reads = float(ntags_ctl1) / float(ntags_ctl2)
if ratio_ctl_reads < 1:
ratio_ctl_reads = 1 / ratio_ctl_reads
ratio_cutoff = 1.2
if ratio_ctl_reads > ratio_cutoff:
logger.info(
"Number of reads in controls differ by > factor of %f. Using pooled controls."
% (ratio_cutoff))
rep1_control = pooled_controls
rep2_control = pooled_controls
else:
if ntags_ctl1 < ntags_rep1:
logger.info(
"Fewer reads in control replicate 1 than experiment replicate 1. Using pooled controls for replicate 1."
)
rep1_control = pooled_controls
rep1_ctl_msg = "pooled controls"
elif not simplicate_experiment and ntags_ctl2 < ntags_rep2:
logger.info(
"Fewer reads in control replicate 2 than experiment replicate 2. Using pooled controls for replicate 2."
)
rep2_control = pooled_controls
rep2_ctl_msg = "pooled controls"
else:
logger.info("Using distinct controls for replicate 1 and 2.")
rep1_control = ctl1_ta # default. May be changed later.
rep2_control = ctl2_ta # default. May be changed later.
rep1_ctl_msg = "control rep1"
rep2_ctl_msg = "control rep2"
common_args = {
'chrom_sizes': chrom_sizes,
'spp_version': spp_version,
'as_file': as_file,
'spp_instance': spp_instance
}
if fragment_length is not None:
common_args.update({'fragment_length': fragment_length})
rep1_peaks_subjob = spp(rep1_ta,
rep1_control,
rep1_xcor,
bigbed=True,
name='Rep1 peaks vs %s' % (rep1_ctl_msg),
prefix='R1',
**common_args)
if not simplicate_experiment:
rep2_peaks_subjob = spp(rep2_ta,
rep2_control,
rep2_xcor,
bigbed=True,
name='Rep2 peaks vs %s' % (rep2_ctl_msg),
prefix='R2',
**common_args)
pooled_peaks_subjob = spp(
pooled_replicates,
control_for_pool,
pooled_replicates_xcor_subjob.get_output_ref("CC_scores_file"),
bigbed=True,
name='Pooled peaks vs %s' % (pool_ctl_msg),
prefix='PL',
**common_args)
output = {
'rep1_peaks': rep1_peaks_subjob.get_output_ref("peaks"),
'rep1_peaks_bb': rep1_peaks_subjob.get_output_ref("peaks_bb"),
'rep1_xcor_plot': rep1_peaks_subjob.get_output_ref("xcor_plot"),
'rep1_xcor_scores': rep1_peaks_subjob.get_output_ref("xcor_scores")
}
if not simplicate_experiment:
output.update({
'rep2_peaks':
rep2_peaks_subjob.get_output_ref("peaks"),
'rep2_peaks_bb':
rep2_peaks_subjob.get_output_ref("peaks_bb"),
'rep2_xcor_plot':
rep2_peaks_subjob.get_output_ref("xcor_plot"),
'rep2_xcor_scores':
rep2_peaks_subjob.get_output_ref("xcor_scores"),
'pooled_peaks':
pooled_peaks_subjob.get_output_ref("peaks"),
'pooled_peaks_bb':
pooled_peaks_subjob.get_output_ref("peaks_bb"),
'pooled_xcor_plot':
pooled_peaks_subjob.get_output_ref("xcor_plot"),
'pooled_xcor_scores':
pooled_peaks_subjob.get_output_ref("xcor_scores")
})
if idr_peaks: # also call peaks on pseudoreplicates for IDR
pseudoreplicator_applet = \
dxpy.find_one_data_object(
classname='applet',
name='pseudoreplicator',
project=dxpy.PROJECT_CONTEXT_ID,
zero_ok=False,
more_ok=False,
return_handler=True)
rep1_pr_subjob = \
pseudoreplicator_applet.run(
{"input_tags": rep1_ta,
"prefix": 'R1PR'},
name='Pseudoreplicate rep1 -> R1PR1,2')
rep1pr1_peaks_subjob = spp(
rep1_pr_subjob.get_output_ref("pseudoreplicate1"),
rep1_control,
rep1_xcor,
bigbed=False,
name='R1PR1 peaks vs %s' % (rep1_ctl_msg),
prefix='R1PR1',
**common_args)
rep1pr2_peaks_subjob = spp(
rep1_pr_subjob.get_output_ref("pseudoreplicate2"),
rep1_control,
rep1_xcor,
bigbed=False,
name='R1PR2 peaks vs %s' % (rep1_ctl_msg),
prefix='R1PR2',
**common_args)
output.update({
'rep1pr1_peaks':
rep1pr1_peaks_subjob.get_output_ref("peaks"),
'rep1pr2_peaks':
rep1pr2_peaks_subjob.get_output_ref("peaks")
})
if not simplicate_experiment:
rep2_pr_subjob = \
pseudoreplicator_applet.run(
{"input_tags": rep2_ta,
"prefix": 'R2PR'},
name='Pseudoreplicate rep2 -> R2PR1,2')
pool_pr1_subjob = pool_applet.run(
{
"inputs": [
rep1_pr_subjob.get_output_ref("pseudoreplicate1"),
rep2_pr_subjob.get_output_ref("pseudoreplicate1")
],
"prefix":
'PPR1'
},
name='Pool R1PR1+R2PR1 -> PPR1')
pool_pr2_subjob = pool_applet.run(
{
"inputs": [
rep1_pr_subjob.get_output_ref("pseudoreplicate2"),
rep2_pr_subjob.get_output_ref("pseudoreplicate2")
],
"prefix":
'PPR2'
},
name='Pool R1PR2+R2PR2 -> PPR2')
rep2pr1_peaks_subjob = spp(
rep2_pr_subjob.get_output_ref("pseudoreplicate1"),
rep2_control,
rep2_xcor,
bigbed=False,
name='R2PR1 peaks vs %s' % (rep2_ctl_msg),
prefix='R2PR1',
**common_args)
rep2pr2_peaks_subjob = spp(
rep2_pr_subjob.get_output_ref("pseudoreplicate2"),
rep2_control,
rep2_xcor,
bigbed=False,
name='R2PR2 peaks vs %s' % (rep2_ctl_msg),
prefix='R2PR2',
**common_args)
pooledpr1_peaks_subjob = spp(
pool_pr1_subjob.get_output_ref("pooled"),
control_for_pool,
pooled_replicates_xcor_subjob.get_output_ref("CC_scores_file"),
bigbed=False,
name='PPR1 peaks vs %s' % (pool_ctl_msg),
prefix='PPR1',
**common_args)
pooledpr2_peaks_subjob = spp(
pool_pr2_subjob.get_output_ref("pooled"),
control_for_pool,
pooled_replicates_xcor_subjob.get_output_ref("CC_scores_file"),
bigbed=False,
name='PPR2 peaks vs %s' % (pool_ctl_msg),
prefix='PPR2',
**common_args)
output.update({
'rep2pr1_peaks':
rep2pr1_peaks_subjob.get_output_ref("peaks"),
'rep2pr2_peaks':
rep2pr2_peaks_subjob.get_output_ref("peaks"),
'pooledpr1_peaks':
pooledpr1_peaks_subjob.get_output_ref("peaks"),
'pooledpr2_peaks':
pooledpr2_peaks_subjob.get_output_ref("peaks"),
})
return output
def main(rep1_ta,
ctl1_ta,
rep1_xcor,
rep1_paired_end,
chrom_sizes,
genomesize,
narrowpeak_as,
gappedpeak_as,
broadpeak_as,
rep2_ta=None,
ctl2_ta=None,
rep2_xcor=None,
rep2_paired_end=None,
fragment_length=None):
rep1_ta_file = rep1_ta
rep1_ta_filename = rep1_ta_file
ntags_rep1 = common.count_lines(rep1_ta_filename)
#
simplicate_experiment = rep1_ta and not rep2_ta
if simplicate_experiment:
logger.info(
"No rep2 tags specified so processing as a simplicate experiment.")
else:
logger.info(
"Rep1 and rep2 tags specified so processing as a replicated experiment."
)
#
if not simplicate_experiment:
assert rep1_paired_end == rep2_paired_end, 'Mixed PE/SE not supported'
rep2_ta_file = rep2_ta
rep2_ta_filename = rep2_ta_file
ntags_rep2 = common.count_lines(rep2_ta_filename)
paired_end = rep1_paired_end
#
unary_control = (ctl1_ta == ctl2_ta) or not ctl2_ta
ctl1_ta_file = ctl1_ta
ctl1_ta_filename = ctl1_ta_file
#
if not unary_control:
ctl2_ta_file = ctl2_ta
ctl2_ta_filename = ctl2_ta_file
else:
ctl2_ta_file = ctl1_ta_file
ctl2_ta_filename = ctl1_ta_file
#
ntags_ctl1 = common.count_lines(ctl1_ta_filename)
ntags_ctl2 = common.count_lines(ctl2_ta_filename)
rep1_control = ctl1_ta # default. May be changed later.
rep1_ctl_msg = "control rep1"
rep2_control = ctl2_ta # default. May be changed later.
rep2_ctl_msg = "control rep2"
#
rep_info = [(ntags_rep1, 'replicate 1', rep1_ta_filename)]
if not simplicate_experiment:
rep_info.append((ntags_rep2, 'replicate 2', rep2_ta_filename))
rep_info.extend([(ntags_ctl1, 'control 1', ctl1_ta_filename),
(ntags_ctl2, 'control 2', ctl2_ta_filename)])
for n, name, filename in rep_info:
logger.info("Found %d tags in %s file %s" % (n, name, filename))
#
subprocess.check_output('ls -l', shell=True, stderr=subprocess.STDOUT)
#
if not simplicate_experiment:
#Pool replicates
pool_replicates_subjob = pool(inputs=[rep1_ta, rep2_ta],
prefix='pooled_reps')
pooled_replicates = pool_replicates_subjob.get("pooled")
#Pool cross-correlation
pooled_replicates_xcor_subjob = xcor_only(pooled_replicates,
paired_end)
#
if unary_control:
logger.info("Only one control supplied.")
if not simplicate_experiment:
logger.info(
"Using one control for both replicate 1 and 2 and for the pool."
)
rep2_control = rep1_control
control_for_pool = rep1_control
pool_ctl_msg = "one control"
else:
#Pool controls
pool_controls_subjob = pool(inputs=[ctl1_ta, ctl2_ta],
prefix="PL_ctls")
pooled_controls = pool_controls_subjob.get("pooled")
# always use the pooled controls for the pool
control_for_pool = pooled_controls
pool_ctl_msg = "pooled controls"
# use the pooled controls for the reps depending on the ratio of rep to
# control reads
ratio_ctl_reads = float(ntags_ctl1) / float(ntags_ctl2)
if ratio_ctl_reads < 1:
ratio_ctl_reads = 1 / ratio_ctl_reads
ratio_cutoff = 1.2
if ratio_ctl_reads > ratio_cutoff:
logger.info(
"Number of reads in controls differ by > factor of %f. Using pooled controls."
% (ratio_cutoff))
rep1_control = pooled_controls
rep2_control = pooled_controls
else:
if ntags_ctl1 < ntags_rep1:
logger.info(
"Fewer reads in control replicate 1 than experiment replicate 1. Using pooled controls for replicate 1."
)
rep1_control = pooled_controls
rep1_ctl_msg = "pooled controls"
elif not simplicate_experiment and ntags_ctl2 < ntags_rep2:
logger.info(
"Fewer reads in control replicate 2 than experiment replicate 2. Using pooled controls for replicate 2."
)
rep2_control = pooled_controls
rep2_ctl_msg = "pooled controls"
else:
logger.info("Using distinct controls for replicate 1 and 2.")
rep1_control = ctl1_ta # default. May be changed later.
rep2_control = ctl2_ta # default. May be changed later.
rep1_ctl_msg = "control rep1"
rep2_ctl_msg = "control rep2"
#
rep1_pr_subjob = pseudoreplicator(input_tags=rep1_ta)
if not simplicate_experiment:
rep2_pr_subjob = pseudoreplicator(input_tags=rep2_ta)
#
pool_pr1_subjob = pool(inputs=[
rep1_pr_subjob.get("pseudoreplicate1"),
rep2_pr_subjob.get("pseudoreplicate1")
],
prefix='PPR1')
pool_pr2_subjob = pool(inputs=[
rep1_pr_subjob.get("pseudoreplicate2"),
rep2_pr_subjob.get("pseudoreplicate2")
],
prefix='PPR2')
#
common_args = {
'chrom_sizes': chrom_sizes,
'genomesize': genomesize,
'narrowpeak_as': narrowpeak_as,
'gappedpeak_as': gappedpeak_as,
'broadpeak_as': broadpeak_as
}
# if the fragment_length argument is given, update macs2 input
if fragment_length is not None:
common_args.update({'fragment_length': fragment_length})
#macs2(experiment, control, xcor_scores_input, chrom_sizes,narrowpeak_as, gappedpeak_as, broadpeak_as, genomesize, prefix=None,fragment_length=None)
common_args.update({'prefix': 'r1'})
rep1_peaks_subjob = macs2(rep1_ta, rep1_control, rep1_xcor, **common_args)
#
common_args.update({'prefix': 'r1pr1'})
rep1pr1_peaks_subjob = macs2(rep1_pr_subjob.get("pseudoreplicate1"),
rep1_control, rep1_xcor, **common_args)
#
common_args.update({'prefix': 'r1pr2'})
rep1pr2_peaks_subjob = macs2(rep1_pr_subjob.get("pseudoreplicate2"),
rep1_control, rep1_xcor, **common_args)
#
if not simplicate_experiment:
common_args.update({'prefix': 'r2'})
rep2_peaks_subjob = macs2(rep2_ta, rep2_control, rep2_xcor,
**common_args)
#
common_args.update({'prefix': 'r2pr1'})
rep2pr1_peaks_subjob = macs2(rep2_pr_subjob.get("pseudoreplicate1"),
rep2_control, rep2_xcor, **common_args)
#
common_args.update({'prefix': 'r2pr2'})
rep2pr2_peaks_subjob = macs2(rep2_pr_subjob.get("pseudoreplicate2"),
rep2_control, rep2_xcor, **common_args)
#
common_args.update({'prefix': 'pool'})
pooled_peaks_subjob = macs2(
pooled_replicates, control_for_pool,
pooled_replicates_xcor_subjob.get("CC_scores_file"), **common_args)
#
common_args.update({'prefix': 'ppr1'})
pooledpr1_peaks_subjob = macs2(
pool_pr1_subjob.get("pooled"), control_for_pool,
pooled_replicates_xcor_subjob.get("CC_scores_file"), **common_args)
#
common_args.update({'prefix': 'ppr2'})
pooledpr2_peaks_subjob = macs2(
pool_pr2_subjob.get("pooled"), control_for_pool,
pooled_replicates_xcor_subjob.get("CC_scores_file"), **common_args)
#
output = {
'rep1_narrowpeaks': rep1_peaks_subjob.get("narrowpeaks"),
'rep1_gappedpeaks': rep1_peaks_subjob.get("gappedpeaks"),
'rep1_broadpeaks': rep1_peaks_subjob.get("broadpeaks"),
'rep1_narrowpeaks_bb': rep1_peaks_subjob.get("narrowpeaks_bb"),
'rep1_gappedpeaks_bb': rep1_peaks_subjob.get("gappedpeaks_bb"),
'rep1_broadpeaks_bb': rep1_peaks_subjob.get("broadpeaks_bb"),
'rep1_fc_signal': rep1_peaks_subjob.get("fc_signal"),
'rep1_pvalue_signal': rep1_peaks_subjob.get("pvalue_signal"),
#
'rep1pr1_narrowpeaks': rep1pr1_peaks_subjob.get("narrowpeaks"),
'rep1pr1_gappedpeaks': rep1pr1_peaks_subjob.get("gappedpeaks"),
'rep1pr1_broadpeaks': rep1pr1_peaks_subjob.get("broadpeaks"),
'rep1pr1_fc_signal': rep1pr1_peaks_subjob.get("fc_signal"),
'rep1pr1_pvalue_signal': rep1pr1_peaks_subjob.get("pvalue_signal"),
#
'rep1pr2_narrowpeaks': rep1pr2_peaks_subjob.get("narrowpeaks"),
'rep1pr2_gappedpeaks': rep1pr2_peaks_subjob.get("gappedpeaks"),
'rep1pr2_broadpeaks': rep1pr2_peaks_subjob.get("broadpeaks"),
'rep1pr2_fc_signal': rep1pr2_peaks_subjob.get("fc_signal"),
'rep1pr2_pvalue_signal': rep1pr2_peaks_subjob.get("pvalue_signal")
}
#
if not simplicate_experiment:
output.update({
'rep2_narrowpeaks':
rep2_peaks_subjob.get("narrowpeaks"),
'rep2_gappedpeaks':
rep2_peaks_subjob.get("gappedpeaks"),
'rep2_broadpeaks':
rep2_peaks_subjob.get("broadpeaks"),
'rep2_narrowpeaks_bb':
rep2_peaks_subjob.get("narrowpeaks_bb"),
'rep2_gappedpeaks_bb':
rep2_peaks_subjob.get("gappedpeaks_bb"),
'rep2_broadpeaks_bb':
rep2_peaks_subjob.get("broadpeaks_bb"),
'rep2_fc_signal':
rep2_peaks_subjob.get("fc_signal"),
'rep2_pvalue_signal':
rep2_peaks_subjob.get("pvalue_signal"),
#
'rep2pr1_narrowpeaks':
rep2pr1_peaks_subjob.get("narrowpeaks"),
'rep2pr1_gappedpeaks':
rep2pr1_peaks_subjob.get("gappedpeaks"),
'rep2pr1_broadpeaks':
rep2pr1_peaks_subjob.get("broadpeaks"),
'rep2pr1_fc_signal':
rep2pr1_peaks_subjob.get("fc_signal"),
'rep2pr1_pvalue_signal':
rep2pr1_peaks_subjob.get("pvalue_signal"),
#
'rep2pr2_narrowpeaks':
rep2pr2_peaks_subjob.get("narrowpeaks"),
'rep2pr2_gappedpeaks':
rep2pr2_peaks_subjob.get("gappedpeaks"),
'rep2pr2_broadpeaks':
rep2pr2_peaks_subjob.get("broadpeaks"),
'rep2pr2_fc_signal':
rep2pr2_peaks_subjob.get("fc_signal"),
'rep2pr2_pvalue_signal':
rep2pr2_peaks_subjob.get("pvalue_signal"),
#
'pooled_narrowpeaks':
pooled_peaks_subjob.get("narrowpeaks"),
'pooled_gappedpeaks':
pooled_peaks_subjob.get("gappedpeaks"),
'pooled_broadpeaks':
pooled_peaks_subjob.get("broadpeaks"),
'pooled_narrowpeaks_bb':
pooled_peaks_subjob.get("narrowpeaks_bb"),
'pooled_gappedpeaks_bb':
pooled_peaks_subjob.get("gappedpeaks_bb"),
'pooled_broadpeaks_bb':
pooled_peaks_subjob.get("broadpeaks_bb"),
'pooled_fc_signal':
pooled_peaks_subjob.get("fc_signal"),
'pooled_pvalue_signal':
pooled_peaks_subjob.get("pvalue_signal"),
#
'pooledpr1_narrowpeaks':
pooledpr1_peaks_subjob.get("narrowpeaks"),
'pooledpr1_gappedpeaks':
pooledpr1_peaks_subjob.get("gappedpeaks"),
'pooledpr1_broadpeaks':
pooledpr1_peaks_subjob.get("broadpeaks"),
'pooledpr1_fc_signal':
pooledpr1_peaks_subjob.get("fc_signal"),
'pooledpr1_pvalue_signal':
pooledpr1_peaks_subjob.get("pvalue_signal"),
#
'pooledpr2_narrowpeaks':
pooledpr2_peaks_subjob.get("narrowpeaks"),
'pooledpr2_gappedpeaks':
pooledpr2_peaks_subjob.get("gappedpeaks"),
'pooledpr2_broadpeaks':
pooledpr2_peaks_subjob.get("broadpeaks"),
'pooledpr2_fc_signal':
pooledpr2_peaks_subjob.get("fc_signal"),
'pooledpr2_pvalue_signal':
pooledpr2_peaks_subjob.get("pvalue_signal")
})
peaks_dirname = '%s_%s_peaks_macs' % (rep1_ta_filename.split(
"/")[-1].split(".")[0], ctl1_ta_filename.split("/")[-1].split(".")[0])
prefix = rep1_ta_filename.split("/")[-1]
peak_file = "%s/%s.peaksfile" % (peaks_dirname, prefix)
with open(peak_file, "w") as fh:
for key, val in output.items():
if isinstance(val, list):
fh.write(": ".join([key, ", ".join(val)]) + "\n")
else:
fh.write(": ".join([key, str(val)]) + "\n")
return output
def internal_pseudoreplicate_IDR(experiment, r1pr_peaks, rep1_ta, rep1_xcor,
paired_end, chrom_sizes, as_file, blacklist,
rep1_signal, fragment_length=None):
r1pr_peaks_file = dxpy.DXFile(r1pr_peaks)
rep1_ta = dxpy.DXFile(rep1_ta)
chrom_sizes_file = dxpy.DXFile(chrom_sizes)
as_file_file = dxpy.DXFile(as_file)
if blacklist is not None:
blacklist_file = dxpy.DXFile(blacklist)
blacklist_filename = 'blacklist_%s' % (blacklist_file.name)
dxpy.download_dxfile(blacklist_file.get_id(), blacklist_filename)
blacklist_filename = common.uncompress(blacklist_filename)
# Need to prepend something to ensure the local filenames will be unique
r1pr_peaks_filename = 'r1pr_%s' % (r1pr_peaks_file.name)
rep1_ta_filename = 'r1ta_%s' % (rep1_ta.name)
chrom_sizes_filename = chrom_sizes_file.name
as_file_filename = as_file_file.name
dxpy.download_dxfile(r1pr_peaks_file.get_id(), r1pr_peaks_filename)
dxpy.download_dxfile(rep1_ta.get_id(), rep1_ta_filename)
dxpy.download_dxfile(chrom_sizes_file.get_id(), chrom_sizes_filename)
dxpy.download_dxfile(as_file_file.get_id(), as_file_filename)
# If fragment_length is given, override appropriate values.
# Calculate, or set the actually used fragment length value.
# Set the fragment_length_given_by_user flag appropriately.
if fragment_length is not None:
rep1_xcor_filename = None
fragment_length_used_rep1 = fragment_length
fragment_length_given_by_user = True
else:
rep1_xcor = dxpy.DXFile(rep1_xcor)
rep1_xcor_filename = 'r1xc_%s' % (rep1_xcor.name)
dxpy.download_dxfile(rep1_xcor.get_id(), rep1_xcor_filename)
fragment_length_used_rep1 = common.xcor_fraglen(rep1_xcor_filename)
fragment_length_given_by_user = False
subprocess.check_output('set -x; ls -l', shell=True)
r1pr_peaks_filename = common.uncompress(r1pr_peaks_filename)
N1 = common.count_lines(r1pr_peaks_filename)
logger.info("%d peaks from rep1 self-pseudoreplicates (N1)" % (N1))
stable_set_filename = "%s_stable.narrowPeak" % (experiment)
if blacklist is not None:
blacklist_filter(r1pr_peaks_filename, stable_set_filename,
blacklist_filename)
Nsb = common.count_lines(stable_set_filename)
logger.info(
"%d peaks blacklisted from the stable set" % (N1-Nsb))
else:
subprocess.check_output(shlex.split(
'cp %s %s' % (r1pr_peaks_filename, stable_set_filename)))
Nsb = N1
logger.info("No blacklist filter applied to the stable set")
# calculate FRiP
n_reads, n_reads_in_peaks, frip_score = common.frip(
rep1_ta_filename, rep1_xcor_filename, stable_set_filename,
chrom_sizes_filename, fragment_length_used_rep1)
output = {
"rep1_frip_nreads": n_reads,
"rep1_frip_nreads_in_peaks": n_reads_in_peaks,
"F1": frip_score,
"fragment_length_used_rep1": fragment_length_used_rep1,
"fragment_length_given_by_user": fragment_length_given_by_user
}
# These are optional outputs to see what's being removed by the blacklist
if blacklist:
output.update({
"pre_bl_stable_set":
dxpy.dxlink(dxpy.upload_local_file(common.compress(
r1pr_peaks_filename)))}
)
# bedtobigbed often fails, so skip creating the bb if it does
stable_set_bb_filename = \
common.bed2bb(stable_set_filename, chrom_sizes_filename,
as_file_filename)
if stable_set_bb_filename:
stable_set_bb_output = \
dxpy.upload_local_file(stable_set_bb_filename)
output.update(
{"stable_set_bb": dxpy.dxlink(stable_set_bb_output)})
output.update({
"N1": N1,
"stable_set": dxpy.dxlink(dxpy.upload_local_file(common.compress(stable_set_filename))),
"Ns": Nsb
})
# These are just passed through for convenience so that signals and tracks
# are available in one place. Both input and output are optional.
if rep1_signal:
output.update({"rep1_signal": rep1_signal})
return output
def main(experiment, control, xcor_scores_input, npeaks, nodups, bigbed, chrom_sizes, as_file=None):
# The following line(s) initialize your data object inputs on the platform
# into dxpy.DXDataObject instances that you can start using immediately.
experiment_file = dxpy.DXFile(experiment)
control_file = dxpy.DXFile(control)
xcor_scores_input_file = dxpy.DXFile(xcor_scores_input)
chrom_sizes_file = dxpy.DXFile(chrom_sizes)
chrom_sizes_filename = chrom_sizes_file.name
dxpy.download_dxfile(chrom_sizes_file.get_id(), chrom_sizes_filename)
if bigbed:
as_file_file = dxpy.DXFile(as_file)
as_file_filename = as_file_file.name
dxpy.download_dxfile(as_file_file.get_id(), as_file_filename)
# The following line(s) download your file inputs to the local file system
# using variable names for the filenames.
experiment_filename = experiment_file.name
dxpy.download_dxfile(experiment_file.get_id(), experiment_filename)
control_filename = control_file.name
dxpy.download_dxfile(control_file.get_id(), control_filename)
xcor_scores_input_filename = xcor_scores_input_file.name
dxpy.download_dxfile(xcor_scores_input_file.get_id(), xcor_scores_input_filename)
output_filename_prefix = experiment_filename.rstrip(".gz").rstrip(".tagAlign")
peaks_filename = output_filename_prefix + ".regionPeak"
final_peaks_filename = peaks_filename + ".gz" # spp adds .gz, so this is the file name that's actually created
xcor_plot_filename = output_filename_prefix + ".pdf"
xcor_scores_filename = output_filename_prefix + ".ccscores"
print subprocess.check_output("ls -l", shell=True, stderr=subprocess.STDOUT)
fraglen_column = 3 # third column in the cross-correlation scores input file
with open(xcor_scores_input_filename, "r") as f:
line = f.readline()
fragment_length = int(line.split("\t")[fraglen_column - 1])
print "Read fragment length: %d" % (fragment_length)
# run_spp_command = subprocess.check_output('which run_spp.R', shell=True)
spp_tarball = "/phantompeakqualtools/spp_1.10.1.tar.gz"
if nodups:
run_spp = "/phantompeakqualtools/run_spp_nodups.R"
else:
run_spp = "/phantompeakqualtools/run_spp.R"
# install spp
print subprocess.check_output("ls -l", shell=True, stderr=subprocess.STDOUT)
print subprocess.check_output(shlex.split("R CMD INSTALL %s" % (spp_tarball)), stderr=subprocess.STDOUT)
spp_command = "Rscript %s -p=%d -c=%s -i=%s -npeak=%d -speak=%d -savr=%s -savp=%s -rf -out=%s" % (
run_spp,
cpu_count(),
experiment_filename,
control_filename,
npeaks,
fragment_length,
peaks_filename,
xcor_plot_filename,
xcor_scores_filename,
)
print spp_command
process = subprocess.Popen(shlex.split(spp_command), stderr=subprocess.STDOUT, stdout=subprocess.PIPE)
for line in iter(process.stdout.readline, ""):
sys.stdout.write(line)
# when one of the peak coordinates are an exact multiple of 10, spp (R) outputs the coordinate in scientific notation
# this changes any such coodinates to decimal notation
# this assumes 10-column output and that the 2nd and 3rd columns are coordinates
# slopBed adjusts feature end coordinates that go off the end of the chromosome
# bedClip removes any features that are still not within the boundaries of the chromosome
fix_coordinate_peaks_filename = output_filename_prefix + ".fixcoord.regionPeak"
out, err = common.run_pipe(
[
"gzip -dc %s" % (final_peaks_filename),
"tee %s" % (peaks_filename),
r"""awk 'BEGIN{OFS="\t"}{print $1,sprintf("%i",$2),sprintf("%i",$3),$4,$5,$6,$7,$8,$9,$10}'""",
"slopBed -i stdin -g %s -b 0" % (chrom_sizes_filename),
"bedClip stdin %s %s" % (chrom_sizes_filename, fix_coordinate_peaks_filename),
]
)
# These lines transfer the peaks files to the temporary workspace for debugging later
# Only at the end are the final files uploaded that will be returned from the applet
dxpy.upload_local_file(peaks_filename)
dxpy.upload_local_file(fix_coordinate_peaks_filename)
n_spp_peaks = common.count_lines(peaks_filename)
print "%s peaks called by spp" % (n_spp_peaks)
print "%s of those peaks removed due to bad coordinates" % (
n_spp_peaks - common.count_lines(fix_coordinate_peaks_filename)
)
print "First 50 peaks"
print subprocess.check_output("head -50 %s" % (fix_coordinate_peaks_filename), shell=True, stderr=subprocess.STDOUT)
if bigbed:
peaks_bb_filename = common.bed2bb(fix_coordinate_peaks_filename, chrom_sizes_filename, as_file_filename)
if peaks_bb_filename:
peaks_bb = dxpy.upload_local_file(peaks_bb_filename)
if not filecmp.cmp(peaks_filename, fix_coordinate_peaks_filename):
print "Returning peaks with fixed coordinates"
print subprocess.check_output(shlex.split("gzip %s" % (fix_coordinate_peaks_filename)))
final_peaks_filename = fix_coordinate_peaks_filename + ".gz"
print subprocess.check_output("ls -l", shell=True, stderr=subprocess.STDOUT)
# print subprocess.check_output('head %s' %(final_peaks_filename), shell=True, stderr=subprocess.STDOUT)
# print subprocess.check_output('head %s' %(xcor_scores_filename), shell=True, stderr=subprocess.STDOUT)
peaks = dxpy.upload_local_file(final_peaks_filename)
xcor_plot = dxpy.upload_local_file(xcor_plot_filename)
xcor_scores = dxpy.upload_local_file(xcor_scores_filename)
output = {}
output["peaks"] = dxpy.dxlink(peaks)
output["xcor_plot"] = dxpy.dxlink(xcor_plot)
output["xcor_scores"] = dxpy.dxlink(xcor_scores)
if bigbed and peaks_bb_filename:
output["peaks_bb"] = dxpy.dxlink(peaks_bb)
return output
def main(rep1_peaks, rep2_peaks, pooled_peaks, pooledpr1_peaks, pooledpr2_peaks,
chrom_sizes, as_file, peak_type, prefix=None,
rep1_signal=None, rep2_signal=None, pooled_signal=None):
# Initialize data object inputs on the platform
# into dxpy.DXDataObject instances
rep1_peaks = dxpy.DXFile(rep1_peaks)
rep2_peaks = dxpy.DXFile(rep2_peaks)
pooled_peaks = dxpy.DXFile(pooled_peaks)
pooledpr1_peaks = dxpy.DXFile(pooledpr1_peaks)
pooledpr2_peaks = dxpy.DXFile(pooledpr2_peaks)
chrom_sizes = dxpy.DXFile(chrom_sizes)
as_file = dxpy.DXFile(as_file)
#Input filenames - necessary to define each explicitly because input files could have the same name, in which case subsequent
#file would overwrite previous file
rep1_peaks_fn = 'rep1-%s' %(rep1_peaks.name)
rep2_peaks_fn = 'rep2-%s' %(rep2_peaks.name)
pooled_peaks_fn = 'pooled-%s' %(pooled_peaks.name)
pooledpr1_peaks_fn = 'pooledpr1-%s' %(pooledpr1_peaks.name)
pooledpr2_peaks_fn = 'pooledpr2-%s' %(pooledpr2_peaks.name)
chrom_sizes_fn = 'chrom.sizes'
as_file_fn = '%s.as' %(peak_type)
# Output filenames
if prefix:
basename = prefix
else:
m = re.match('(.*)(\.%s)+(\.((gz)|(Z)|(bz)|(bz2)))' %(peak_type), pooled_peaks.name) #strip off the peak and compression extensions
if m:
basename = m.group(1)
else:
basename = pooled_peaks.name
overlapping_peaks_fn = '%s.replicated.%s' %(basename, peak_type)
overlapping_peaks_bb_fn = overlapping_peaks_fn + '.bb'
rejected_peaks_fn = '%s.rejected.%s' %(basename, peak_type)
rejected_peaks_bb_fn = rejected_peaks_fn + '.bb'
# Intermediate filenames
overlap_tr_fn = 'replicated_tr.%s' %(peak_type)
overlap_pr_fn = 'replicated_pr.%s' %(peak_type)
# Download file inputs to the local file system with local filenames
dxpy.download_dxfile(rep1_peaks.get_id(), rep1_peaks_fn)
dxpy.download_dxfile(rep2_peaks.get_id(), rep2_peaks_fn)
dxpy.download_dxfile(pooled_peaks.get_id(), pooled_peaks_fn)
dxpy.download_dxfile(pooledpr1_peaks.get_id(), pooledpr1_peaks_fn)
dxpy.download_dxfile(pooledpr2_peaks.get_id(), pooledpr2_peaks_fn)
dxpy.download_dxfile(chrom_sizes.get_id(), chrom_sizes_fn)
dxpy.download_dxfile(as_file.get_id(), as_file_fn)
'''
#find pooled peaks that are in (rep1 AND rep2)
out, err = common.run_pipe([
'intersectBed -wa -f 0.50 -r -a %s -b %s' %(pooled_peaks_fn, rep1_peaks_fn),
'intersectBed -wa -f 0.50 -r -a stdin -b %s' %(rep2_peaks_fn)
], overlap_tr_fn)
print "%d peaks overlap with both true replicates" %(common.count_lines(overlap_tr_fn))
#pooled peaks that are in (pooledpseudorep1 AND pooledpseudorep2)
out, err = common.run_pipe([
'intersectBed -wa -f 0.50 -r -a %s -b %s' %(pooled_peaks_fn, pooledpr1_peaks_fn),
'intersectBed -wa -f 0.50 -r -a stdin -b %s' %(pooledpr2_peaks_fn)
], overlap_pr_fn)
print "%d peaks overlap with both pooled pseudoreplicates" %(common.count_lines(overlap_pr_fn))
#combined pooled peaks in (rep1 AND rep2) OR (pooledpseudorep1 AND pooledpseudorep2)
out, err = common.run_pipe([
'intersectBed -wa -a %s -b %s %s' %(pooled_peaks_fn, overlap_tr_fn, overlap_pr_fn),
'intersectBed -wa -u -a %s -b stdin' %(pooled_peaks_fn)
], overlapping_peaks_fn)
print "%d peaks overall with true replicates or with pooled pseudorepliates" %(common.count_lines(overlapping_peaks_fn))
'''
#the only difference between the peak_types is how the extra columns are handled
if peak_type == "narrowPeak":
awk_command = r"""awk 'BEGIN{FS="\t";OFS="\t"}{s1=$3-$2; s2=$13-$12; if (($21/s1 >= 0.5) || ($21/s2 >= 0.5)) {print $0}}'"""
cut_command = 'cut -f 1-10'
bed_type = 'bed6+4'
elif peak_type == "gappedPeak":
awk_command = r"""awk 'BEGIN{FS="\t";OFS="\t"}{s1=$3-$2; s2=$18-$17; if (($31/s1 >= 0.5) || ($31/s2 >= 0.5)) {print $0}}'"""
cut_command = 'cut -f 1-15'
bed_type = 'bed12+3'
elif peak_type == "broadPeak":
awk_command = r"""awk 'BEGIN{FS="\t";OFS="\t"}{s1=$3-$2; s2=$12-$11; if (($19/s1 >= 0.5) || ($19/s2 >= 0.5)) {print $0}}'"""
cut_command = 'cut -f 1-9'
bed_type = 'bed6+3'
else:
assert peak_type in ['narrowPeak', 'gappedPeak', 'broadPeak'], "%s is unrecognized. peak_type should be narrowPeak, gappedPeak or broadPeak." % (peak_type)
# Find pooled peaks that overlap Rep1 and Rep2 where overlap is defined as the fractional overlap wrt any one of the overlapping peak pairs > 0.5
out, err = common.run_pipe([
'intersectBed -wo -a %s -b %s' %(pooled_peaks_fn, rep1_peaks_fn),
awk_command,
cut_command,
'sort -u',
'intersectBed -wo -a stdin -b %s' %(rep2_peaks_fn),
awk_command,
cut_command,
'sort -u'
], overlap_tr_fn)
print "%d peaks overlap with both true replicates" %(common.count_lines(overlap_tr_fn))
# Find pooled peaks that overlap PseudoRep1 and PseudoRep2 where overlap is defined as the fractional overlap wrt any one of the overlapping peak pairs > 0.5
out, err = common.run_pipe([
'intersectBed -wo -a %s -b %s' %(pooled_peaks_fn, pooledpr1_peaks_fn),
awk_command,
cut_command,
'sort -u',
'intersectBed -wo -a stdin -b %s' %(pooledpr2_peaks_fn),
awk_command,
cut_command,
'sort -u'
], overlap_pr_fn)
print "%d peaks overlap with both pooled pseudoreplicates" %(common.count_lines(overlap_pr_fn))
# Combine peak lists
out, err = common.run_pipe([
'cat %s %s' %(overlap_tr_fn, overlap_pr_fn),
'sort -u'
], overlapping_peaks_fn)
print "%d peaks overlap with true replicates or with pooled pseudorepliates" %(common.count_lines(overlapping_peaks_fn))
#rejected peaks
out, err = common.run_pipe([
'intersectBed -wa -v -a %s -b %s' %(pooled_peaks_fn, overlapping_peaks_fn)
], rejected_peaks_fn)
print "%d peaks were rejected" %(common.count_lines(rejected_peaks_fn))
npeaks_in = common.count_lines(common.uncompress(pooled_peaks_fn))
npeaks_out = common.count_lines(overlapping_peaks_fn)
npeaks_rejected = common.count_lines(rejected_peaks_fn)
#make bigBed files for visualization
overlapping_peaks_bb_fn = common.bed2bb(overlapping_peaks_fn, chrom_sizes_fn, as_file_fn, bed_type=bed_type)
rejected_peaks_bb_fn = common.bed2bb(rejected_peaks_fn, chrom_sizes_fn, as_file_fn, bed_type=bed_type)
# overlapping_peaks_bb_fn = common.bed2bb(common.slop_clip(overlapping_peaks_fn, chrom_sizes_fn, "gappedPeak"), chrom_sizes_fn, as_file_fn, bed_type=bed_type)
# rejected_peaks_bb_fn = common.bed2bb(common.slop_clip(rejected_peaks_fn, chrom_sizes_fn, "gappedPeak"), chrom_sizes_fn, as_file_fn, bed_type=bed_type)
# Upload file outputs from the local file system.
overlapping_peaks = dxpy.upload_local_file(common.compress(overlapping_peaks_fn))
overlapping_peaks_bb = dxpy.upload_local_file(overlapping_peaks_bb_fn)
rejected_peaks = dxpy.upload_local_file(common.compress(rejected_peaks_fn))
rejected_peaks_bb = dxpy.upload_local_file(rejected_peaks_bb_fn)
# The following line fills in some basic dummy output and assumes
# that you have created variables to represent your output with
# the same name as your output fields.
output = {
"overlapping_peaks" : dxpy.dxlink(overlapping_peaks),
"overlapping_peaks_bb" : dxpy.dxlink(overlapping_peaks_bb),
"rejected_peaks" : dxpy.dxlink(rejected_peaks),
"rejected_peaks_bb" : dxpy.dxlink(rejected_peaks_bb),
"npeaks_in" : npeaks_in,
"npeaks_out" : npeaks_out,
'npeaks_rejected' : npeaks_rejected
}
# These are just passed through for convenience so that signals and tracks
# are available in one place. Both input and output are optional.
if rep1_signal:
output.update({"rep1_signal": rep1_signal})
if rep2_signal:
output.update({"rep2_signal": rep2_signal})
if pooled_signal:
output.update({"pooled_signal": pooled_signal})
return output
def internal_pseudoreplicate_IDR(experiment,
r1pr_peaks,
rep1_ta,
rep1_xcor,
paired_end,
chrom_sizes,
as_file,
blacklist,
rep1_signal,
fragment_length=None):
r1pr_peaks_file = dxpy.DXFile(r1pr_peaks)
rep1_ta = dxpy.DXFile(rep1_ta)
chrom_sizes_file = dxpy.DXFile(chrom_sizes)
as_file_file = dxpy.DXFile(as_file)
if blacklist is not None:
blacklist_file = dxpy.DXFile(blacklist)
blacklist_filename = 'blacklist_%s' % (blacklist_file.name)
dxpy.download_dxfile(blacklist_file.get_id(), blacklist_filename)
blacklist_filename = common.uncompress(blacklist_filename)
# Need to prepend something to ensure the local filenames will be unique
r1pr_peaks_filename = 'r1pr_%s' % (r1pr_peaks_file.name)
rep1_ta_filename = 'r1ta_%s' % (rep1_ta.name)
chrom_sizes_filename = chrom_sizes_file.name
as_file_filename = as_file_file.name
dxpy.download_dxfile(r1pr_peaks_file.get_id(), r1pr_peaks_filename)
dxpy.download_dxfile(rep1_ta.get_id(), rep1_ta_filename)
dxpy.download_dxfile(chrom_sizes_file.get_id(), chrom_sizes_filename)
dxpy.download_dxfile(as_file_file.get_id(), as_file_filename)
# If fragment_length is given, override appropriate values.
# Calculate, or set the actually used fragment length value.
# Set the fragment_length_given_by_user flag appropriately.
if fragment_length is not None:
rep1_xcor_filename = None
fragment_length_used_rep1 = fragment_length
fragment_length_given_by_user = True
else:
rep1_xcor = dxpy.DXFile(rep1_xcor)
rep1_xcor_filename = 'r1xc_%s' % (rep1_xcor.name)
dxpy.download_dxfile(rep1_xcor.get_id(), rep1_xcor_filename)
fragment_length_used_rep1 = common.xcor_fraglen(rep1_xcor_filename)
fragment_length_given_by_user = False
subprocess.check_output('set -x; ls -l', shell=True)
r1pr_peaks_filename = common.uncompress(r1pr_peaks_filename)
N1 = common.count_lines(r1pr_peaks_filename)
logger.info("%d peaks from rep1 self-pseudoreplicates (N1)" % (N1))
stable_set_filename = "%s_stable.narrowPeak" % (experiment)
if blacklist is not None:
blacklist_filter(r1pr_peaks_filename, stable_set_filename,
blacklist_filename)
Nsb = common.count_lines(stable_set_filename)
logger.info("%d peaks blacklisted from the stable set" % (N1 - Nsb))
else:
subprocess.check_output(
shlex.split('cp %s %s' %
(r1pr_peaks_filename, stable_set_filename)))
Nsb = N1
logger.info("No blacklist filter applied to the stable set")
# calculate FRiP
n_reads, n_reads_in_peaks, frip_score = common.frip(
rep1_ta_filename, rep1_xcor_filename, stable_set_filename,
chrom_sizes_filename, fragment_length_used_rep1)
output = {
"rep1_frip_nreads": n_reads,
"rep1_frip_nreads_in_peaks": n_reads_in_peaks,
"F1": frip_score,
"fragment_length_used_rep1": fragment_length_used_rep1,
"fragment_length_given_by_user": fragment_length_given_by_user
}
# These are optional outputs to see what's being removed by the blacklist
if blacklist:
output.update({
"pre_bl_stable_set":
dxpy.dxlink(
dxpy.upload_local_file(common.compress(r1pr_peaks_filename)))
})
# bedtobigbed often fails, so skip creating the bb if it does
stable_set_bb_filename = \
common.bed2bb(stable_set_filename, chrom_sizes_filename,
as_file_filename)
if stable_set_bb_filename:
stable_set_bb_output = \
dxpy.upload_local_file(stable_set_bb_filename)
output.update({"stable_set_bb": dxpy.dxlink(stable_set_bb_output)})
output.update({
"N1":
N1,
"stable_set":
dxpy.dxlink(
dxpy.upload_local_file(common.compress(stable_set_filename))),
"Ns":
Nsb
})
# These are just passed through for convenience so that signals and tracks
# are available in one place. Both input and output are optional.
if rep1_signal:
output.update({"rep1_signal": rep1_signal})
return output
def replicated_IDR(experiment,
reps_peaks,
r1pr_peaks,
r2pr_peaks,
pooledpr_peaks,
rep1_ta,
rep1_xcor,
rep2_ta,
rep2_xcor,
paired_end,
chrom_sizes,
as_file,
blacklist,
rep1_signal,
rep2_signal,
pooled_signal,
fragment_length=None):
# TODO for now just taking the peak files. This applet should actually
# call IDR instead of putting that in the workflow populator script
reps_peaks_file = dxpy.DXFile(reps_peaks)
r1pr_peaks_file = dxpy.DXFile(r1pr_peaks)
r2pr_peaks_file = dxpy.DXFile(r2pr_peaks)
pooledpr_peaks_file = dxpy.DXFile(pooledpr_peaks)
rep1_ta_file = dxpy.DXFile(rep1_ta)
rep2_ta_file = dxpy.DXFile(rep2_ta)
rep1_xcor_file = dxpy.DXFile(rep1_xcor)
rep2_xcor_file = dxpy.DXFile(rep2_xcor)
chrom_sizes_file = dxpy.DXFile(chrom_sizes)
as_file_file = dxpy.DXFile(as_file)
if blacklist is not None:
blacklist_file = dxpy.DXFile(blacklist)
blacklist_filename = 'blacklist_%s' % (blacklist_file.name)
dxpy.download_dxfile(blacklist_file.get_id(), blacklist_filename)
blacklist_filename = common.uncompress(blacklist_filename)
# Need to prepend something to ensure the local filenames will be unique
reps_peaks_filename = 'true_%s' % (reps_peaks_file.name)
r1pr_peaks_filename = 'r1pr_%s' % (r1pr_peaks_file.name)
r2pr_peaks_filename = 'r2pr_%s' % (r2pr_peaks_file.name)
pooledpr_peaks_filename = 'pooledpr_%s' % (pooledpr_peaks_file.name)
rep1_ta_filename = 'r1ta_%s' % (rep1_ta_file.name)
rep2_ta_filename = 'r2ta_%s' % (rep2_ta_file.name)
rep1_xcor_filename = 'r1cc_%s' % (rep1_xcor_file.name)
rep2_xcor_filename = 'r2cc_%s' % (rep2_xcor_file.name)
chrom_sizes_filename = chrom_sizes_file.name
as_file_filename = as_file_file.name
dxpy.download_dxfile(reps_peaks_file.get_id(), reps_peaks_filename)
dxpy.download_dxfile(r1pr_peaks_file.get_id(), r1pr_peaks_filename)
dxpy.download_dxfile(r2pr_peaks_file.get_id(), r2pr_peaks_filename)
dxpy.download_dxfile(pooledpr_peaks_file.get_id(), pooledpr_peaks_filename)
dxpy.download_dxfile(rep1_ta_file.get_id(), rep1_ta_filename)
dxpy.download_dxfile(rep2_ta_file.get_id(), rep2_ta_filename)
dxpy.download_dxfile(rep1_xcor_file.get_id(), rep1_xcor_filename)
dxpy.download_dxfile(rep2_xcor_file.get_id(), rep2_xcor_filename)
dxpy.download_dxfile(chrom_sizes_file.get_id(), chrom_sizes_filename)
dxpy.download_dxfile(as_file_file.get_id(), as_file_filename)
reps_peaks_filename = common.uncompress(reps_peaks_filename)
r1pr_peaks_filename = common.uncompress(r1pr_peaks_filename)
r2pr_peaks_filename = common.uncompress(r2pr_peaks_filename)
pooledpr_peaks_filename = common.uncompress(pooledpr_peaks_filename)
pool_applet = dxpy.find_one_data_object(classname='applet',
name='pool',
project=dxpy.PROJECT_CONTEXT_ID,
zero_ok=False,
more_ok=False,
return_handler=True)
pool_replicates_subjob = \
pool_applet.run(
{"inputs": [rep1_ta, rep2_ta],
"prefix": 'pooled_reps'},
name='Pool replicates')
# next call could be on 267 and save time?
pool_replicates_subjob.wait_on_done()
# If fragment_length is not given, calculate the fragment_length
# using crosscorrelation. Else use the overridevalue. Set the
# pool_xcor_filename to None to accommodate common.frip calls.
# Calculate, or set, actually used fragment lengths for different
# cases. Set the flag indicating whether the fragment length
# was given by the user.
if fragment_length is not None:
pool_xcor_filename = None
fragment_length_used_rep1 = fragment_length
fragment_length_used_rep2 = fragment_length
fragment_length_used_pool = fragment_length
fragment_length_given_by_user = True
else:
pooled_replicates_xcor_subjob = \
xcor_only(
pool_replicates_subjob.get_output_ref("pooled"),
paired_end,
spp_version=None,
name='Pool cross-correlation')
pooled_replicates_xcor_subjob.wait_on_done()
pool_xcor_link = pooled_replicates_xcor_subjob.describe(
)['output'].get("CC_scores_file")
pool_xcor_file = dxpy.get_handler(pool_xcor_link)
pool_xcor_filename = 'poolcc_%s' % (pool_xcor_file.name)
dxpy.download_dxfile(pool_xcor_file.get_id(), pool_xcor_filename)
fragment_length_used_rep1 = common.xcor_fraglen(rep1_xcor_filename)
fragment_length_used_rep2 = common.xcor_fraglen(rep2_xcor_filename)
fragment_length_used_pool = common.xcor_fraglen(pool_xcor_filename)
fragment_length_given_by_user = False
pool_ta_link = pool_replicates_subjob.describe()['output'].get("pooled")
pool_ta_file = dxpy.get_handler(pool_ta_link)
pool_ta_filename = 'poolta_%s' % (pool_ta_file.name)
dxpy.download_dxfile(pool_ta_file.get_id(), pool_ta_filename)
logger.info(subprocess.check_output('set -x; ls -l', shell=True))
Nt = common.count_lines(reps_peaks_filename)
logger.info("%d peaks from true replicates (Nt)" % (Nt))
N1 = common.count_lines(r1pr_peaks_filename)
logger.info("%d peaks from rep1 self-pseudoreplicates (N1)" % (N1))
N2 = common.count_lines(r2pr_peaks_filename)
logger.info("%d peaks from rep2 self-pseudoreplicates (N2)" % (N2))
Np = common.count_lines(pooledpr_peaks_filename)
logger.info("%d peaks from pooled pseudoreplicates (Np)" % (Np))
# generate the conservative set, which is always based on the IDR peaks
# from true replicates
conservative_set_filename = \
'%s_final_conservative.narrowPeak' % (experiment)
if blacklist is not None:
blacklist_filter(reps_peaks_filename, conservative_set_filename,
blacklist_filename)
Ncb = common.count_lines(conservative_set_filename)
logger.info("%d peaks blacklisted from the conservative set" %
(Nt - Ncb))
else:
subprocess.check_output(
shlex.split('cp %s %s' %
(reps_peaks_filename, conservative_set_filename)))
Ncb = Nt
logger.info("No blacklist filter applied to the conservative set")
# generate the optimal set, which is based on the longest of IDR peaks
# list from true reps or the IDR peaks from the pseudoreplicates of the
# pool
if Nt >= Np:
peaks_to_filter_filename = reps_peaks_filename
No = Nt
else:
peaks_to_filter_filename = pooledpr_peaks_filename
No = Np
optimal_set_filename = '%s_final_optimal.narrowPeak' % (experiment)
if blacklist is not None:
blacklist_filter(peaks_to_filter_filename, optimal_set_filename,
blacklist_filename)
Nob = common.count_lines(optimal_set_filename)
logger.info("%d peaks blacklisted from the optimal set" % (No - Nob))
else:
subprocess.check_output(
shlex.split('cp %s %s' %
(peaks_to_filter_filename, optimal_set_filename)))
Nob = No
logger.info("No blacklist filter applied to the optimal set")
rescue_ratio = float(max(Np, Nt)) / float(min(Np, Nt))
self_consistency_ratio = float(max(N1, N2)) / float(min(N1, N2))
if rescue_ratio > 2 and self_consistency_ratio > 2:
reproducibility = 'fail'
elif rescue_ratio > 2 or self_consistency_ratio > 2:
reproducibility = 'borderline'
else:
reproducibility = 'pass'
# FRiP (fraction reads in peaks)
# rep1 stable peaks comparing internal pseudoreplicates
rep1_n_reads, rep1_n_reads_in_peaks, rep1_frip_score = common.frip(
rep1_ta_filename, rep1_xcor_filename, r1pr_peaks_filename,
chrom_sizes_filename, fragment_length)
# rep2 stable peaks comparing internal pseudoreplicates
rep2_n_reads, rep2_n_reads_in_peaks, rep2_frip_score = common.frip(
rep2_ta_filename, rep2_xcor_filename, r2pr_peaks_filename,
chrom_sizes_filename, fragment_length)
# comparing true reps
true_n_reads, true_n_reads_in_peaks, true_frip_score = common.frip(
pool_ta_filename, pool_xcor_filename, reps_peaks_filename,
chrom_sizes_filename, fragment_length)
# comparing pooled pseudoreplicates
pr_n_reads, pr_n_reads_in_peaks, pr_frip_score = common.frip(
pool_ta_filename, pool_xcor_filename, pooledpr_peaks_filename,
chrom_sizes_filename, fragment_length)
output = {
"rep1_frip_nreads": rep1_n_reads,
"rep1_frip_nreads_in_peaks": rep1_n_reads_in_peaks,
"F1": rep1_frip_score,
"rep2_frip_nreads": rep2_n_reads,
"rep2_frip_nreads_in_peaks": rep2_n_reads_in_peaks,
"F2": rep2_frip_score,
"true_frip_nreads": true_n_reads,
"true_frip_nreads_in_peaks": true_n_reads_in_peaks,
"Ft": true_frip_score,
"pr_frip_nreads": pr_n_reads,
"pr_frip_nreads_in_peaks": pr_n_reads_in_peaks,
"Fp": pr_frip_score,
"fragment_length_used_rep1": fragment_length_used_rep1,
"fragment_length_used_rep2": fragment_length_used_rep2,
"fragment_length_used_pool": fragment_length_used_pool,
"fragment_length_given_by_user": fragment_length_given_by_user
}
# These are optional outputs to see what's being removed by the blacklist
if blacklist:
output.update({
"pre_bl_conservative_set":
dxpy.dxlink(
dxpy.upload_local_file(common.compress(reps_peaks_filename))),
"pre_bl_optimal_set":
dxpy.dxlink(
dxpy.upload_local_file(
common.compress(peaks_to_filter_filename)))
})
# bedtobigbed often fails, so skip creating the bb if it does
conservative_set_bb_filename = \
common.bed2bb(conservative_set_filename, chrom_sizes_filename,
as_file_filename)
optimal_set_bb_filename = \
common.bed2bb(optimal_set_filename, chrom_sizes_filename,
as_file_filename)
if conservative_set_bb_filename:
conservative_set_bb_output = \
dxpy.upload_local_file(conservative_set_bb_filename)
output.update(
{"conservative_set_bb": dxpy.dxlink(conservative_set_bb_output)})
if optimal_set_bb_filename:
optimal_set_bb_output = dxpy.upload_local_file(optimal_set_bb_filename)
output.update({"optimal_set_bb": dxpy.dxlink(optimal_set_bb_output)})
output.update({
"Nt":
Nt,
"N1":
N1,
"N2":
N2,
"Np":
Np,
"conservative_set":
dxpy.dxlink(
dxpy.upload_local_file(
common.compress(conservative_set_filename))),
"optimal_set":
dxpy.dxlink(
dxpy.upload_local_file(common.compress(optimal_set_filename))),
"rescue_ratio":
rescue_ratio,
"self_consistency_ratio":
self_consistency_ratio,
"reproducibility_test":
reproducibility,
"No":
Nob,
"Nc":
Ncb
})
# These are just passed through for convenience so that signals and tracks
# are available in one place. Both input and output are optional.
if rep1_signal:
output.update({"rep1_signal": rep1_signal})
if rep2_signal:
output.update({"rep2_signal": rep2_signal})
if pooled_signal:
output.update({"pooled_signal": pooled_signal})
return output
def main(rep1_ta, ctl1_ta, rep1_paired_end,
rep2_ta=None, ctl2_ta=None, rep2_paired_end=None):
rep1_ta_filename = rep1_ta
ntags_rep1 = common.count_lines(rep1_ta_filename)
output = {'rep1_ta': rep1_ta_filename}
simplicate_experiment = rep1_ta and not rep2_ta
output.update({'simplicate_experiment': simplicate_experiment})
if simplicate_experiment:
logger.info("No rep2 tags specified so processing as a simplicate experiment.")
else:
logger.info("Rep1 and rep2 tags specified so processing as a replicated experiment.")
output.update({'rep2_ta': rep2_ta})
if not simplicate_experiment:
assert rep1_paired_end == rep2_paired_end, 'Mixed PE/SE not supported'
rep2_ta_filename = rep2_ta
ntags_rep2 = common.count_lines(rep2_ta_filename)
output.update({'rep2_ta': rep2_ta_filename})
paired_end = rep1_paired_end
output.update({'paired_end': paired_end})
unary_control = (ctl1_ta == ctl2_ta) or not ctl2_ta
ctl1_ta_filename = ctl1_ta
if not unary_control:
ctl2_ta_filename = ctl2_ta
else:
ctl2_ta_filename = ctl1_ta
ntags_ctl1 = common.count_lines(ctl1_ta_filename)
ntags_ctl2 = common.count_lines(ctl2_ta_filename)
rep1_control = ctl1_ta # default. May be changed later.
rep2_control = ctl2_ta # default. May be changed later.
output.update({'rep1_control': rep1_control,
'rep2_control': rep2_control})
rep_info = [(ntags_rep1, 'replicate 1', rep1_ta_filename)]
if not simplicate_experiment:
rep_info.append((ntags_rep2, 'replicate 2', rep2_ta_filename))
rep_info.extend(
[(ntags_ctl1, 'control 1', ctl1_ta_filename),
(ntags_ctl2, 'control 2', ctl2_ta_filename)])
for n, name, filename in rep_info:
logger.info("Found %d tags in %s file %s" % (n, name, filename))
subprocess.check_output('ls -l', shell=True, stderr=subprocess.STDOUT)
if not simplicate_experiment:
pool_replicates_subjob = \
pool(**{"inputs": [rep1_ta, rep2_ta],
"prefix": 'pooled_reps'})
pooled_replicates = pool_replicates_subjob.get("pooled")
output.update({'pooled_replicates': pooled_replicates})
# this needs to go to the other image
'''
pooled_replicates_xcor_subjob = \
xcor_only(
pooled_replicates,
paired_end,
name='Pool cross-correlation')
'''
if unary_control:
logger.info("Only one control supplied.")
if not simplicate_experiment:
logger.info("Using one control for both replicate 1 and 2 and for the pool.")
rep2_control = rep1_control
control_for_pool = rep1_control
output.update({'rep2_control': rep2_control,
'control_for_pool': rep1_control})
else:
pool_controls_subjob = pool(
**{"inputs": [ctl1_ta, ctl2_ta],
"prefix": "PL_ctls"})
pooled_controls = pool_controls_subjob.get("pooled")
# always use the pooled controls for the pool
control_for_pool = pooled_controls
output.update({'control_for_pool': control_for_pool})
# use the pooled controls for the reps depending on the ratio of rep to
# control reads
ratio_ctl_reads = float(ntags_ctl1)/float(ntags_ctl2)
if ratio_ctl_reads < 1:
ratio_ctl_reads = 1/ratio_ctl_reads
ratio_cutoff = 1.2
if ratio_ctl_reads > ratio_cutoff:
logger.info(
"Number of reads in controls differ by > factor of %f. Using pooled controls."
% (ratio_cutoff))
rep1_control = pooled_controls
rep2_control = pooled_controls
output.update({'rep1_control': pooled_controls,
'rep2_control': pooled_controls})
else:
if ntags_ctl1 < ntags_rep1:
logger.info("Fewer reads in control replicate 1 than experiment replicate 1. Using pooled controls for replicate 1.")
rep1_control = pooled_controls
output.update({'rep1_control': pooled_controls})
elif not simplicate_experiment and ntags_ctl2 < ntags_rep2:
logger.info("Fewer reads in control replicate 2 than experiment replicate 2. Using pooled controls for replicate 2.")
rep2_control = pooled_controls
output.update({'rep2_control': pooled_controls})
else:
logger.info(
"Using distinct controls for replicate 1 and 2.")
rep1_control = ctl1_ta
rep2_control = ctl2_ta
output.update({'rep1_control': ctl1_ta,
'rep2_control': ctl2_ta})
rep1_pr_subjob = pseudoreplicator(**{"input_tags": rep1_ta})
r1pr1 = rep1_pr_subjob.get('pseudoreplicate1')
r1pr2 = rep1_pr_subjob.get('pseudoreplicate2')
output.update({'r1pr1': r1pr1,
'r1pr2': r1pr2})
if not simplicate_experiment:
rep2_pr_subjob = pseudoreplicator(**{"input_tags": rep2_ta})
r2pr1 = rep2_pr_subjob.get('pseudoreplicate1')
r2pr2 = rep2_pr_subjob.get('pseudoreplicate2')
output.update({'r2pr1': r2pr1,
'r2pr2': r2pr2})
pool_pr1_subjob = pool(
**{"inputs": [rep1_pr_subjob.get("pseudoreplicate1"),
rep2_pr_subjob.get("pseudoreplicate1")],
"prefix": 'PPR1'})
pool_pr2_subjob = pool(
**{"inputs": [rep1_pr_subjob.get("pseudoreplicate2"),
rep2_pr_subjob.get("pseudoreplicate2")],
"prefix": 'PPR2'})
ppr1 = pool_pr1_subjob.get('pooled')
ppr2 = pool_pr2_subjob.get('pooled')
output.update({'ppr1': ppr1,
'ppr2': ppr2})
# should there be an indication of the simplicateness of the
# experiment in the output json? this could be a good way to
# direct the next step without putting too much logic into the
# workflow. ADDED.
# Turns out Cromwell does not support reading .json. Instead
# they have read_map function that accepts 2 column TSVs.
with open('pool_and_pseudoreplicate_outfiles.mapping', 'w') as f:
for key in output:
f.write('%s\t%s\n' % (key, output[key]))
return output
def main(rep1_peaks,
rep2_peaks,
pooled_peaks,
pooledpr1_peaks,
pooledpr2_peaks,
chrom_sizes,
as_file,
peak_type,
prefix=None,
rep1_signal=None,
rep2_signal=None,
pooled_signal=None):
# Initialize data object inputs on the platform
# into dxpy.DXDataObject instances
rep1_peaks = dxpy.DXFile(rep1_peaks)
rep2_peaks = dxpy.DXFile(rep2_peaks)
pooled_peaks = dxpy.DXFile(pooled_peaks)
pooledpr1_peaks = dxpy.DXFile(pooledpr1_peaks)
pooledpr2_peaks = dxpy.DXFile(pooledpr2_peaks)
chrom_sizes = dxpy.DXFile(chrom_sizes)
as_file = dxpy.DXFile(as_file)
#Input filenames - necessary to define each explicitly because input files could have the same name, in which case subsequent
#file would overwrite previous file
rep1_peaks_fn = 'rep1-%s' % (rep1_peaks.name)
rep2_peaks_fn = 'rep2-%s' % (rep2_peaks.name)
pooled_peaks_fn = 'pooled-%s' % (pooled_peaks.name)
pooledpr1_peaks_fn = 'pooledpr1-%s' % (pooledpr1_peaks.name)
pooledpr2_peaks_fn = 'pooledpr2-%s' % (pooledpr2_peaks.name)
chrom_sizes_fn = 'chrom.sizes'
as_file_fn = '%s.as' % (peak_type)
# Output filenames
if prefix:
basename = prefix
else:
m = re.match(
'(.*)(\.%s)+(\.((gz)|(Z)|(bz)|(bz2)))' % (peak_type),
pooled_peaks.name) #strip off the peak and compression extensions
if m:
basename = m.group(1)
else:
basename = pooled_peaks.name
overlapping_peaks_fn = '%s.replicated.%s' % (basename, peak_type)
overlapping_peaks_bb_fn = overlapping_peaks_fn + '.bb'
rejected_peaks_fn = '%s.rejected.%s' % (basename, peak_type)
rejected_peaks_bb_fn = rejected_peaks_fn + '.bb'
# Intermediate filenames
overlap_tr_fn = 'replicated_tr.%s' % (peak_type)
overlap_pr_fn = 'replicated_pr.%s' % (peak_type)
# Download file inputs to the local file system with local filenames
dxpy.download_dxfile(rep1_peaks.get_id(), rep1_peaks_fn)
dxpy.download_dxfile(rep2_peaks.get_id(), rep2_peaks_fn)
dxpy.download_dxfile(pooled_peaks.get_id(), pooled_peaks_fn)
dxpy.download_dxfile(pooledpr1_peaks.get_id(), pooledpr1_peaks_fn)
dxpy.download_dxfile(pooledpr2_peaks.get_id(), pooledpr2_peaks_fn)
dxpy.download_dxfile(chrom_sizes.get_id(), chrom_sizes_fn)
dxpy.download_dxfile(as_file.get_id(), as_file_fn)
'''
#find pooled peaks that are in (rep1 AND rep2)
out, err = common.run_pipe([
'intersectBed -wa -f 0.50 -r -a %s -b %s' %(pooled_peaks_fn, rep1_peaks_fn),
'intersectBed -wa -f 0.50 -r -a stdin -b %s' %(rep2_peaks_fn)
], overlap_tr_fn)
print "%d peaks overlap with both true replicates" %(common.count_lines(overlap_tr_fn))
#pooled peaks that are in (pooledpseudorep1 AND pooledpseudorep2)
out, err = common.run_pipe([
'intersectBed -wa -f 0.50 -r -a %s -b %s' %(pooled_peaks_fn, pooledpr1_peaks_fn),
'intersectBed -wa -f 0.50 -r -a stdin -b %s' %(pooledpr2_peaks_fn)
], overlap_pr_fn)
print "%d peaks overlap with both pooled pseudoreplicates" %(common.count_lines(overlap_pr_fn))
#combined pooled peaks in (rep1 AND rep2) OR (pooledpseudorep1 AND pooledpseudorep2)
out, err = common.run_pipe([
'intersectBed -wa -a %s -b %s %s' %(pooled_peaks_fn, overlap_tr_fn, overlap_pr_fn),
'intersectBed -wa -u -a %s -b stdin' %(pooled_peaks_fn)
], overlapping_peaks_fn)
print "%d peaks overall with true replicates or with pooled pseudorepliates" %(common.count_lines(overlapping_peaks_fn))
'''
#the only difference between the peak_types is how the extra columns are handled
if peak_type == "narrowPeak":
awk_command = r"""awk 'BEGIN{FS="\t";OFS="\t"}{s1=$3-$2; s2=$13-$12; if (($21/s1 >= 0.5) || ($21/s2 >= 0.5)) {print $0}}'"""
cut_command = 'cut -f 1-10'
bed_type = 'bed6+4'
elif peak_type == "gappedPeak":
awk_command = r"""awk 'BEGIN{FS="\t";OFS="\t"}{s1=$3-$2; s2=$18-$17; if (($31/s1 >= 0.5) || ($31/s2 >= 0.5)) {print $0}}'"""
cut_command = 'cut -f 1-15'
bed_type = 'bed12+3'
elif peak_type == "broadPeak":
awk_command = r"""awk 'BEGIN{FS="\t";OFS="\t"}{s1=$3-$2; s2=$12-$11; if (($19/s1 >= 0.5) || ($19/s2 >= 0.5)) {print $0}}'"""
cut_command = 'cut -f 1-9'
bed_type = 'bed6+3'
else:
assert peak_type in [
'narrowPeak', 'gappedPeak', 'broadPeak'
], "%s is unrecognized. peak_type should be narrowPeak, gappedPeak or broadPeak." % (
peak_type)
# Find pooled peaks that overlap Rep1 and Rep2 where overlap is defined as the fractional overlap wrt any one of the overlapping peak pairs > 0.5
out, err = common.run_pipe([
'intersectBed -wo -a %s -b %s' %
(pooled_peaks_fn, rep1_peaks_fn), awk_command, cut_command, 'sort -u',
'intersectBed -wo -a stdin -b %s' %
(rep2_peaks_fn), awk_command, cut_command, 'sort -u'
], overlap_tr_fn)
print "%d peaks overlap with both true replicates" % (
common.count_lines(overlap_tr_fn))
# Find pooled peaks that overlap PseudoRep1 and PseudoRep2 where overlap is defined as the fractional overlap wrt any one of the overlapping peak pairs > 0.5
out, err = common.run_pipe([
'intersectBed -wo -a %s -b %s' % (pooled_peaks_fn, pooledpr1_peaks_fn),
awk_command, cut_command, 'sort -u',
'intersectBed -wo -a stdin -b %s' %
(pooledpr2_peaks_fn), awk_command, cut_command, 'sort -u'
], overlap_pr_fn)
print "%d peaks overlap with both pooled pseudoreplicates" % (
common.count_lines(overlap_pr_fn))
# Combine peak lists
out, err = common.run_pipe(
['cat %s %s' % (overlap_tr_fn, overlap_pr_fn), 'sort -u'],
overlapping_peaks_fn)
print "%d peaks overlap with true replicates or with pooled pseudorepliates" % (
common.count_lines(overlapping_peaks_fn))
#rejected peaks
out, err = common.run_pipe([
'intersectBed -wa -v -a %s -b %s' %
(pooled_peaks_fn, overlapping_peaks_fn)
], rejected_peaks_fn)
print "%d peaks were rejected" % (common.count_lines(rejected_peaks_fn))
npeaks_in = common.count_lines(common.uncompress(pooled_peaks_fn))
npeaks_out = common.count_lines(overlapping_peaks_fn)
npeaks_rejected = common.count_lines(rejected_peaks_fn)
#make bigBed files for visualization
overlapping_peaks_bb_fn = common.bed2bb(overlapping_peaks_fn,
chrom_sizes_fn,
as_file_fn,
bed_type=bed_type)
rejected_peaks_bb_fn = common.bed2bb(rejected_peaks_fn,
chrom_sizes_fn,
as_file_fn,
bed_type=bed_type)
# overlapping_peaks_bb_fn = common.bed2bb(common.slop_clip(overlapping_peaks_fn, chrom_sizes_fn, "gappedPeak"), chrom_sizes_fn, as_file_fn, bed_type=bed_type)
# rejected_peaks_bb_fn = common.bed2bb(common.slop_clip(rejected_peaks_fn, chrom_sizes_fn, "gappedPeak"), chrom_sizes_fn, as_file_fn, bed_type=bed_type)
# Upload file outputs from the local file system.
overlapping_peaks = dxpy.upload_local_file(
common.compress(overlapping_peaks_fn))
overlapping_peaks_bb = dxpy.upload_local_file(overlapping_peaks_bb_fn)
rejected_peaks = dxpy.upload_local_file(common.compress(rejected_peaks_fn))
rejected_peaks_bb = dxpy.upload_local_file(rejected_peaks_bb_fn)
# The following line fills in some basic dummy output and assumes
# that you have created variables to represent your output with
# the same name as your output fields.
output = {
"overlapping_peaks": dxpy.dxlink(overlapping_peaks),
"overlapping_peaks_bb": dxpy.dxlink(overlapping_peaks_bb),
"rejected_peaks": dxpy.dxlink(rejected_peaks),
"rejected_peaks_bb": dxpy.dxlink(rejected_peaks_bb),
"npeaks_in": npeaks_in,
"npeaks_out": npeaks_out,
'npeaks_rejected': npeaks_rejected
}
# These are just passed through for convenience so that signals and tracks
# are available in one place. Both input and output are optional.
if rep1_signal:
output.update({"rep1_signal": rep1_signal})
if rep2_signal:
output.update({"rep2_signal": rep2_signal})
if pooled_signal:
output.update({"pooled_signal": pooled_signal})
return output
def main(experiment, reps_peaks, r1pr_peaks, r2pr_peaks, pooledpr_peaks,
chrom_sizes, as_file, blacklist=None,
rep1_signal=None, rep2_signal=None, pooled_signal=None):
# TODO for now just taking the peak files. This applet should actually
# call IDR instead of putting that in the workflow populator script
reps_peaks_file = dxpy.DXFile(reps_peaks)
r1pr_peaks_file = dxpy.DXFile(r1pr_peaks)
r2pr_peaks_file = dxpy.DXFile(r2pr_peaks)
pooledpr_peaks_file = dxpy.DXFile(pooledpr_peaks)
chrom_sizes_file = dxpy.DXFile(chrom_sizes)
as_file_file = dxpy.DXFile(as_file)
if blacklist is not None:
blacklist_file = dxpy.DXFile(blacklist)
blacklist_filename = 'blacklist_%s' % (blacklist_file.name)
dxpy.download_dxfile(blacklist_file.get_id(), blacklist_filename)
blacklist_filename = common.uncompress(blacklist_filename)
# Need to prepend something to ensure the local filenames will be unique
reps_peaks_filename = 'true_%s' % (reps_peaks_file.name)
r1pr_peaks_filename = 'r1pr_%s' % (r1pr_peaks_file.name)
r2pr_peaks_filename = 'r2pr_%s' % (r2pr_peaks_file.name)
pooledpr_peaks_filename = 'pooledpr_%s' % (pooledpr_peaks_file.name)
chrom_sizes_filename = chrom_sizes_file.name
as_file_filename = as_file_file.name
dxpy.download_dxfile(reps_peaks_file.get_id(), reps_peaks_filename)
dxpy.download_dxfile(r1pr_peaks_file.get_id(), r1pr_peaks_filename)
dxpy.download_dxfile(r2pr_peaks_file.get_id(), r2pr_peaks_filename)
dxpy.download_dxfile(pooledpr_peaks_file.get_id(), pooledpr_peaks_filename)
dxpy.download_dxfile(chrom_sizes_file.get_id(), chrom_sizes_filename)
dxpy.download_dxfile(as_file_file.get_id(), as_file_filename)
subprocess.check_output('set -x; ls -l', shell=True)
reps_peaks_filename = common.uncompress(reps_peaks_filename)
r1pr_peaks_filename = common.uncompress(r1pr_peaks_filename)
r2pr_peaks_filename = common.uncompress(r2pr_peaks_filename)
pooledpr_peaks_filename = common.uncompress(pooledpr_peaks_filename)
Nt = common.count_lines(reps_peaks_filename)
logger.info("%d peaks from true replicates (Nt)" % (Nt))
N1 = common.count_lines(r1pr_peaks_filename)
logger.info("%d peaks from rep1 self-pseudoreplicates (N1)" % (N1))
N2 = common.count_lines(r2pr_peaks_filename)
logger.info("%d peaks from rep2 self-pseudoreplicates (N2)" % (N2))
Np = common.count_lines(pooledpr_peaks_filename)
logger.info("%d peaks from pooled pseudoreplicates (Np)" % (Np))
# generate the conservative set, which is always based on the IDR peaks
# from true replicates
conservative_set_filename = \
'%s_final_conservative.narrowPeak' % (experiment)
if blacklist is not None:
blacklist_filter(reps_peaks_filename, conservative_set_filename,
blacklist_filename)
Ncb = common.count_lines(conservative_set_filename)
logger.info(
"%d peaks blacklisted from the conservative set" % (Nt-Ncb))
else:
subprocess.check_output(shlex.split(
'cp %s %s' % (reps_peaks_filename, conservative_set_filename)))
Ncb = Nt
logger.info("No blacklist filter applied to the conservative set")
# generate the optimal set, which is based on the longest of IDR peaks
# list from true reps or the IDR peaks from the pseudoreplicates of the
# pool
if Nt >= Np:
peaks_to_filter_filename = reps_peaks_filename
No = Nt
else:
peaks_to_filter_filename = pooledpr_peaks_filename
No = Np
optimal_set_filename = '%s_final_optimal.narrowPeak' % (experiment)
if blacklist is not None:
blacklist_filter(peaks_to_filter_filename, optimal_set_filename,
blacklist_filename)
Nob = common.count_lines(optimal_set_filename)
logger.info("%d peaks blacklisted from the optimal set" % (No-Nob))
else:
subprocess.check_output(shlex.split(
'cp %s %s' % (peaks_to_filter_filename, optimal_set_filename)))
Nob = No
logger.info("No blacklist filter applied to the optimal set")
rescue_ratio = float(max(Np, Nt)) / float(min(Np, Nt))
self_consistency_ratio = float(max(N1, N2)) / float(min(N1, N2))
if rescue_ratio > 2 and self_consistency_ratio > 2:
reproducibility = 'fail'
elif rescue_ratio > 2 or self_consistency_ratio > 2:
reproducibility = 'borderline'
else:
reproducibility = 'pass'
output = {}
# These are optional outputs to see what's being removed by the blacklist
if blacklist:
output.update({
"pre_bl_conservative_set":
dxpy.dxlink(dxpy.upload_local_file(common.compress(
reps_peaks_filename))),
"pre_bl_optimal_set":
dxpy.dxlink(dxpy.upload_local_file(common.compress(
peaks_to_filter_filename)))}
)
# bedtobigbed often fails, so skip creating the bb if it does
conservative_set_bb_filename = \
common.bed2bb(conservative_set_filename, chrom_sizes_filename,
as_file_filename)
optimal_set_bb_filename = \
common.bed2bb(optimal_set_filename, chrom_sizes_filename,
as_file_filename)
if conservative_set_bb_filename:
conservative_set_bb_output = \
dxpy.upload_local_file(conservative_set_bb_filename)
output.update(
{"conservative_set_bb": dxpy.dxlink(conservative_set_bb_output)})
if optimal_set_bb_filename:
optimal_set_bb_output = dxpy.upload_local_file(optimal_set_bb_filename)
output.update(
{"optimal_set_bb": dxpy.dxlink(optimal_set_bb_output)})
output.update({
"Nt": Nt,
"N1": N1,
"N2": N2,
"Np": Np,
"conservative_set": dxpy.dxlink(dxpy.upload_local_file(common.compress(conservative_set_filename))),
"optimal_set": dxpy.dxlink(dxpy.upload_local_file(common.compress(optimal_set_filename))),
"rescue_ratio": rescue_ratio,
"self_consistency_ratio": self_consistency_ratio,
"reproducibility_test": reproducibility,
"No": Nob,
"Nc": Ncb
})
# These are just passed through for convenience so that signals and tracks
# are available in one place. Both input and output are optional.
if rep1_signal:
output.update({"rep1_signal": rep1_signal})
if rep2_signal:
output.update({"rep2_signal": rep2_signal})
if pooled_signal:
output.update({"pooled_signal": pooled_signal})
logging.info("Exiting with output: %s", output)
return output
def replicated_IDR(experiment,
reps_peaks, r1pr_peaks, r2pr_peaks, pooledpr_peaks,
rep1_ta, rep1_xcor, rep2_ta, rep2_xcor,
paired_end, chrom_sizes, as_file, blacklist,
rep1_signal, rep2_signal, pooled_signal,
fragment_length=None):
# TODO for now just taking the peak files. This applet should actually
# call IDR instead of putting that in the workflow populator script
reps_peaks_file = dxpy.DXFile(reps_peaks)
r1pr_peaks_file = dxpy.DXFile(r1pr_peaks)
r2pr_peaks_file = dxpy.DXFile(r2pr_peaks)
pooledpr_peaks_file = dxpy.DXFile(pooledpr_peaks)
rep1_ta_file = dxpy.DXFile(rep1_ta)
rep2_ta_file = dxpy.DXFile(rep2_ta)
rep1_xcor_file = dxpy.DXFile(rep1_xcor)
rep2_xcor_file = dxpy.DXFile(rep2_xcor)
chrom_sizes_file = dxpy.DXFile(chrom_sizes)
as_file_file = dxpy.DXFile(as_file)
if blacklist is not None:
blacklist_file = dxpy.DXFile(blacklist)
blacklist_filename = 'blacklist_%s' % (blacklist_file.name)
dxpy.download_dxfile(blacklist_file.get_id(), blacklist_filename)
blacklist_filename = common.uncompress(blacklist_filename)
# Need to prepend something to ensure the local filenames will be unique
reps_peaks_filename = 'true_%s' % (reps_peaks_file.name)
r1pr_peaks_filename = 'r1pr_%s' % (r1pr_peaks_file.name)
r2pr_peaks_filename = 'r2pr_%s' % (r2pr_peaks_file.name)
pooledpr_peaks_filename = 'pooledpr_%s' % (pooledpr_peaks_file.name)
rep1_ta_filename = 'r1ta_%s' % (rep1_ta_file.name)
rep2_ta_filename = 'r2ta_%s' % (rep2_ta_file.name)
rep1_xcor_filename = 'r1cc_%s' % (rep1_xcor_file.name)
rep2_xcor_filename = 'r2cc_%s' % (rep2_xcor_file.name)
chrom_sizes_filename = chrom_sizes_file.name
as_file_filename = as_file_file.name
dxpy.download_dxfile(reps_peaks_file.get_id(), reps_peaks_filename)
dxpy.download_dxfile(r1pr_peaks_file.get_id(), r1pr_peaks_filename)
dxpy.download_dxfile(r2pr_peaks_file.get_id(), r2pr_peaks_filename)
dxpy.download_dxfile(pooledpr_peaks_file.get_id(), pooledpr_peaks_filename)
dxpy.download_dxfile(rep1_ta_file.get_id(), rep1_ta_filename)
dxpy.download_dxfile(rep2_ta_file.get_id(), rep2_ta_filename)
dxpy.download_dxfile(rep1_xcor_file.get_id(), rep1_xcor_filename)
dxpy.download_dxfile(rep2_xcor_file.get_id(), rep2_xcor_filename)
dxpy.download_dxfile(chrom_sizes_file.get_id(), chrom_sizes_filename)
dxpy.download_dxfile(as_file_file.get_id(), as_file_filename)
reps_peaks_filename = common.uncompress(reps_peaks_filename)
r1pr_peaks_filename = common.uncompress(r1pr_peaks_filename)
r2pr_peaks_filename = common.uncompress(r2pr_peaks_filename)
pooledpr_peaks_filename = common.uncompress(pooledpr_peaks_filename)
pool_applet = dxpy.find_one_data_object(
classname='applet',
name='pool',
project=dxpy.PROJECT_CONTEXT_ID,
zero_ok=False,
more_ok=False,
return_handler=True)
pool_replicates_subjob = \
pool_applet.run(
{"inputs": [rep1_ta, rep2_ta],
"prefix": 'pooled_reps'},
name='Pool replicates')
# next call could be on 267 and save time?
pool_replicates_subjob.wait_on_done()
# If fragment_length is not given, calculate the fragment_length
# using crosscorrelation. Else use the overridevalue. Set the
# pool_xcor_filename to None to accommodate common.frip calls.
# Calculate, or set, actually used fragment lengths for different
# cases. Set the flag indicating whether the fragment length
# was given by the user.
if fragment_length is not None:
pool_xcor_filename = None
fragment_length_used_rep1 = fragment_length
fragment_length_used_rep2 = fragment_length
fragment_length_used_pool = fragment_length
fragment_length_given_by_user = True
else:
pooled_replicates_xcor_subjob = \
xcor_only(
pool_replicates_subjob.get_output_ref("pooled"),
paired_end,
spp_version=None,
name='Pool cross-correlation')
pooled_replicates_xcor_subjob.wait_on_done()
pool_xcor_link = pooled_replicates_xcor_subjob.describe()['output'].get("CC_scores_file")
pool_xcor_file = dxpy.get_handler(pool_xcor_link)
pool_xcor_filename = 'poolcc_%s' % (pool_xcor_file.name)
dxpy.download_dxfile(pool_xcor_file.get_id(), pool_xcor_filename)
fragment_length_used_rep1 = common.xcor_fraglen(rep1_xcor_filename)
fragment_length_used_rep2 = common.xcor_fraglen(rep2_xcor_filename)
fragment_length_used_pool = common.xcor_fraglen(pool_xcor_filename)
fragment_length_given_by_user = False
pool_ta_link = pool_replicates_subjob.describe()['output'].get("pooled")
pool_ta_file = dxpy.get_handler(pool_ta_link)
pool_ta_filename = 'poolta_%s' % (pool_ta_file.name)
dxpy.download_dxfile(pool_ta_file.get_id(), pool_ta_filename)
logger.info(subprocess.check_output('set -x; ls -l', shell=True))
Nt = common.count_lines(reps_peaks_filename)
logger.info("%d peaks from true replicates (Nt)" % (Nt))
N1 = common.count_lines(r1pr_peaks_filename)
logger.info("%d peaks from rep1 self-pseudoreplicates (N1)" % (N1))
N2 = common.count_lines(r2pr_peaks_filename)
logger.info("%d peaks from rep2 self-pseudoreplicates (N2)" % (N2))
Np = common.count_lines(pooledpr_peaks_filename)
logger.info("%d peaks from pooled pseudoreplicates (Np)" % (Np))
# generate the conservative set, which is always based on the IDR peaks
# from true replicates
conservative_set_filename = \
'%s_final_conservative.narrowPeak' % (experiment)
if blacklist is not None:
blacklist_filter(reps_peaks_filename, conservative_set_filename,
blacklist_filename)
Ncb = common.count_lines(conservative_set_filename)
logger.info(
"%d peaks blacklisted from the conservative set" % (Nt-Ncb))
else:
subprocess.check_output(shlex.split(
'cp %s %s' % (reps_peaks_filename, conservative_set_filename)))
Ncb = Nt
logger.info("No blacklist filter applied to the conservative set")
# generate the optimal set, which is based on the longest of IDR peaks
# list from true reps or the IDR peaks from the pseudoreplicates of the
# pool
if Nt >= Np:
peaks_to_filter_filename = reps_peaks_filename
No = Nt
else:
peaks_to_filter_filename = pooledpr_peaks_filename
No = Np
optimal_set_filename = '%s_final_optimal.narrowPeak' % (experiment)
if blacklist is not None:
blacklist_filter(peaks_to_filter_filename, optimal_set_filename,
blacklist_filename)
Nob = common.count_lines(optimal_set_filename)
logger.info("%d peaks blacklisted from the optimal set" % (No-Nob))
else:
subprocess.check_output(shlex.split(
'cp %s %s' % (peaks_to_filter_filename, optimal_set_filename)))
Nob = No
logger.info("No blacklist filter applied to the optimal set")
rescue_ratio = float(max(Np, Nt)) / float(min(Np, Nt))
self_consistency_ratio = float(max(N1, N2)) / float(min(N1, N2))
if rescue_ratio > 2 and self_consistency_ratio > 2:
reproducibility = 'fail'
elif rescue_ratio > 2 or self_consistency_ratio > 2:
reproducibility = 'borderline'
else:
reproducibility = 'pass'
# FRiP (fraction reads in peaks)
# rep1 stable peaks comparing internal pseudoreplicates
rep1_n_reads, rep1_n_reads_in_peaks, rep1_frip_score = common.frip(
rep1_ta_filename, rep1_xcor_filename, r1pr_peaks_filename,
chrom_sizes_filename, fragment_length)
# rep2 stable peaks comparing internal pseudoreplicates
rep2_n_reads, rep2_n_reads_in_peaks, rep2_frip_score = common.frip(
rep2_ta_filename, rep2_xcor_filename, r2pr_peaks_filename,
chrom_sizes_filename, fragment_length)
# comparing true reps
true_n_reads, true_n_reads_in_peaks, true_frip_score = common.frip(
pool_ta_filename, pool_xcor_filename, reps_peaks_filename,
chrom_sizes_filename, fragment_length)
# comparing pooled pseudoreplicates
pr_n_reads, pr_n_reads_in_peaks, pr_frip_score = common.frip(
pool_ta_filename, pool_xcor_filename, pooledpr_peaks_filename,
chrom_sizes_filename, fragment_length)
output = {
"rep1_frip_nreads" : rep1_n_reads,
"rep1_frip_nreads_in_peaks" : rep1_n_reads_in_peaks,
"F1" : rep1_frip_score,
"rep2_frip_nreads" : rep2_n_reads,
"rep2_frip_nreads_in_peaks" : rep2_n_reads_in_peaks,
"F2" : rep2_frip_score,
"true_frip_nreads" : true_n_reads,
"true_frip_nreads_in_peaks" : true_n_reads_in_peaks,
"Ft" : true_frip_score,
"pr_frip_nreads" : pr_n_reads,
"pr_frip_nreads_in_peaks" : pr_n_reads_in_peaks,
"Fp" : pr_frip_score,
"fragment_length_used_rep1": fragment_length_used_rep1,
"fragment_length_used_rep2": fragment_length_used_rep2,
"fragment_length_used_pool": fragment_length_used_pool,
"fragment_length_given_by_user": fragment_length_given_by_user
}
# These are optional outputs to see what's being removed by the blacklist
if blacklist:
output.update({
"pre_bl_conservative_set":
dxpy.dxlink(dxpy.upload_local_file(common.compress(
reps_peaks_filename))),
"pre_bl_optimal_set":
dxpy.dxlink(dxpy.upload_local_file(common.compress(
peaks_to_filter_filename)))}
)
# bedtobigbed often fails, so skip creating the bb if it does
conservative_set_bb_filename = \
common.bed2bb(conservative_set_filename, chrom_sizes_filename,
as_file_filename)
optimal_set_bb_filename = \
common.bed2bb(optimal_set_filename, chrom_sizes_filename,
as_file_filename)
if conservative_set_bb_filename:
conservative_set_bb_output = \
dxpy.upload_local_file(conservative_set_bb_filename)
output.update(
{"conservative_set_bb": dxpy.dxlink(conservative_set_bb_output)})
if optimal_set_bb_filename:
optimal_set_bb_output = dxpy.upload_local_file(optimal_set_bb_filename)
output.update(
{"optimal_set_bb": dxpy.dxlink(optimal_set_bb_output)})
output.update({
"Nt": Nt,
"N1": N1,
"N2": N2,
"Np": Np,
"conservative_set": dxpy.dxlink(dxpy.upload_local_file(common.compress(conservative_set_filename))),
"optimal_set": dxpy.dxlink(dxpy.upload_local_file(common.compress(optimal_set_filename))),
"rescue_ratio": rescue_ratio,
"self_consistency_ratio": self_consistency_ratio,
"reproducibility_test": reproducibility,
"No": Nob,
"Nc": Ncb
})
# These are just passed through for convenience so that signals and tracks
# are available in one place. Both input and output are optional.
if rep1_signal:
output.update({"rep1_signal": rep1_signal})
if rep2_signal:
output.update({"rep2_signal": rep2_signal})
if pooled_signal:
output.update({"pooled_signal": pooled_signal})
return output
def main(experiment, control, xcor_scores_input, npeaks, nodups, bigbed, chrom_sizes, as_file=None, prefix=None):
# The following line(s) initialize your data object inputs on the platform
# into dxpy.DXDataObject instances that you can start using immediately.
experiment_file = dxpy.DXFile(experiment)
control_file = dxpy.DXFile(control)
xcor_scores_input_file = dxpy.DXFile(xcor_scores_input)
chrom_sizes_file = dxpy.DXFile(chrom_sizes)
chrom_sizes_filename = chrom_sizes_file.name
dxpy.download_dxfile(chrom_sizes_file.get_id(), chrom_sizes_filename)
if bigbed:
as_file_file = dxpy.DXFile(as_file)
as_file_filename = as_file_file.name
dxpy.download_dxfile(as_file_file.get_id(), as_file_filename)
# The following line(s) download your file inputs to the local file system
# using variable names for the filenames.
experiment_filename = experiment_file.name
dxpy.download_dxfile(experiment_file.get_id(), experiment_filename)
control_filename = control_file.name
dxpy.download_dxfile(control_file.get_id(), control_filename)
xcor_scores_input_filename = xcor_scores_input_file.name
dxpy.download_dxfile(xcor_scores_input_file.get_id(), xcor_scores_input_filename)
if not prefix:
output_filename_prefix = experiment_filename.rstrip('.gz').rstrip('.tagAlign')
else:
output_filename_prefix = prefix
peaks_filename = output_filename_prefix + '.regionPeak'
final_peaks_filename = peaks_filename + '.gz' #spp adds .gz, so this is the file name that's actually created
xcor_plot_filename = output_filename_prefix + '.pdf'
xcor_scores_filename = output_filename_prefix + '.ccscores'
print subprocess.check_output('ls -l', shell=True, stderr=subprocess.STDOUT)
fraglen_column = 3 # third column in the cross-correlation scores input file
with open(xcor_scores_input_filename, 'r') as f:
line = f.readline()
fragment_length = int(line.split('\t')[fraglen_column-1])
print "Read fragment length: %d" %(fragment_length)
#run_spp_command = subprocess.check_output('which run_spp.R', shell=True)
spp_tarball = '/phantompeakqualtools/spp_1.10.1.tar.gz'
if nodups:
run_spp = '/phantompeakqualtools/run_spp_nodups.R'
else:
run_spp = '/phantompeakqualtools/run_spp.R'
#install spp
subprocess.check_call('ls -l', shell=True)
subprocess.check_call(shlex.split('R CMD INSTALL %s' %(spp_tarball)))
spp_command = "Rscript %s -p=%d -c=%s -i=%s -npeak=%d -speak=%d -savr=%s -savp=%s -rf -out=%s" %(run_spp, cpu_count(), experiment_filename, control_filename, npeaks, fragment_length, peaks_filename, xcor_plot_filename, xcor_scores_filename)
print spp_command
# process = subprocess.Popen(shlex.split(spp_command), stderr=subprocess.STDOUT, stdout=subprocess.PIPE)
# for line in iter(process.stdout.readline, ''):
# sys.stdout.write(line)
subprocess.check_call(shlex.split(spp_command))
#when one of the peak coordinates are an exact multiple of 10, spp (R) outputs the coordinate in scientific notation
#this changes any such coodinates to decimal notation
#this assumes 10-column output and that the 2nd and 3rd columns are coordinates
#slopBed adjusts feature end coordinates that go off the end of the chromosome
#bedClip removes any features that are still not within the boundaries of the chromosome
fix_coordinate_peaks_filename = output_filename_prefix + '.fixcoord.regionPeak'
out, err = common.run_pipe([
"gzip -dc %s" %(final_peaks_filename),
"tee %s" %(peaks_filename),
r"""awk 'BEGIN{OFS="\t"}{print $1,sprintf("%i",$2),sprintf("%i",$3),$4,$5,$6,$7,$8,$9,$10}'""",
'slopBed -i stdin -g %s -b 0' %(chrom_sizes_filename),
'bedClip stdin %s %s' %(chrom_sizes_filename, fix_coordinate_peaks_filename)
])
#These lines transfer the peaks files to the temporary workspace for debugging later
#Only at the end are the final files uploaded that will be returned from the applet
dxpy.upload_local_file(peaks_filename)
dxpy.upload_local_file(fix_coordinate_peaks_filename)
n_spp_peaks = common.count_lines(peaks_filename)
print "%s peaks called by spp" %(n_spp_peaks)
print "%s of those peaks removed due to bad coordinates" %(n_spp_peaks - common.count_lines(fix_coordinate_peaks_filename))
print "First 50 peaks"
print subprocess.check_output('head -50 %s' %(fix_coordinate_peaks_filename), shell=True, stderr=subprocess.STDOUT)
if bigbed:
peaks_bb_filename = common.bed2bb(fix_coordinate_peaks_filename, chrom_sizes_filename, as_file_filename)
if peaks_bb_filename:
peaks_bb = dxpy.upload_local_file(peaks_bb_filename)
if not filecmp.cmp(peaks_filename,fix_coordinate_peaks_filename):
print "Returning peaks with fixed coordinates"
print subprocess.check_output(shlex.split('gzip %s' %(fix_coordinate_peaks_filename)))
final_peaks_filename = fix_coordinate_peaks_filename + '.gz'
print subprocess.check_output('ls -l', shell=True, stderr=subprocess.STDOUT)
#print subprocess.check_output('head %s' %(final_peaks_filename), shell=True, stderr=subprocess.STDOUT)
#print subprocess.check_output('head %s' %(xcor_scores_filename), shell=True, stderr=subprocess.STDOUT)
peaks = dxpy.upload_local_file(final_peaks_filename)
xcor_plot = dxpy.upload_local_file(xcor_plot_filename)
xcor_scores = dxpy.upload_local_file(xcor_scores_filename)
output = {}
output["peaks"] = dxpy.dxlink(peaks)
output["xcor_plot"] = dxpy.dxlink(xcor_plot)
output["xcor_scores"] = dxpy.dxlink(xcor_scores)
if bigbed and peaks_bb_filename:
output["peaks_bb"] = dxpy.dxlink(peaks_bb)
return output
def main(rep1_ta, ctl1_ta, rep1_xcor, rep1_paired_end, chrom_sizes, genomesize,
narrowpeak_as, gappedpeak_as, broadpeak_as,
rep2_ta=None, ctl2_ta=None, rep2_xcor=None, rep2_paired_end=None,
fragment_length=None):
rep1_ta_file = dxpy.DXFile(rep1_ta)
dxpy.download_dxfile(rep1_ta_file.get_id(), rep1_ta_file.name)
rep1_ta_filename = rep1_ta_file.name
ntags_rep1 = common.count_lines(rep1_ta_filename)
simplicate_experiment = rep1_ta and not rep2_ta
if simplicate_experiment:
logger.info("No rep2 tags specified so processing as a simplicate experiment.")
else:
logger.info("Rep1 and rep2 tags specified so processing as a replicated experiment.")
if not simplicate_experiment:
assert rep1_paired_end == rep2_paired_end, 'Mixed PE/SE not supported'
rep2_ta_file = dxpy.DXFile(rep2_ta)
dxpy.download_dxfile(rep2_ta_file.get_id(), rep2_ta_file.name)
rep2_ta_filename = rep2_ta_file.name
ntags_rep2 = common.count_lines(rep2_ta_filename)
paired_end = rep1_paired_end
unary_control = (ctl1_ta == ctl2_ta) or not ctl2_ta
ctl1_ta_file = dxpy.DXFile(ctl1_ta)
dxpy.download_dxfile(ctl1_ta_file.get_id(), ctl1_ta_file.name)
ctl1_ta_filename = ctl1_ta_file.name
if not unary_control:
ctl2_ta_file = dxpy.DXFile(ctl2_ta)
dxpy.download_dxfile(ctl2_ta_file.get_id(), ctl2_ta_file.name)
ctl2_ta_filename = ctl2_ta_file.name
else:
ctl2_ta_file = ctl1_ta_file
ctl2_ta_filename = ctl1_ta_file.name
ntags_ctl1 = common.count_lines(ctl1_ta_filename)
ntags_ctl2 = common.count_lines(ctl2_ta_filename)
rep1_control = ctl1_ta # default. May be changed later.
rep1_ctl_msg = "control rep1"
rep2_control = ctl2_ta # default. May be changed later.
rep2_ctl_msg = "control rep2"
rep_info = [(ntags_rep1, 'replicate 1', rep1_ta_filename)]
if not simplicate_experiment:
rep_info.append((ntags_rep2, 'replicate 2', rep2_ta_filename))
rep_info.extend(
[(ntags_ctl1, 'control 1', ctl1_ta_filename),
(ntags_ctl2, 'control 2', ctl2_ta_filename)])
for n, name, filename in rep_info:
logger.info("Found %d tags in %s file %s" % (n, name, filename))
subprocess.check_output('ls -l', shell=True, stderr=subprocess.STDOUT)
if not simplicate_experiment:
pool_applet = dxpy.find_one_data_object(
classname='applet',
name='pool',
project=dxpy.PROJECT_CONTEXT_ID,
zero_ok=False,
more_ok=False,
return_handler=True)
pool_replicates_subjob = \
pool_applet.run(
{"inputs": [rep1_ta, rep2_ta],
"prefix": 'pooled_reps'},
name='Pool replicates')
pooled_replicates = pool_replicates_subjob.get_output_ref("pooled")
pooled_replicates_xcor_subjob = \
xcor_only(
pooled_replicates,
paired_end,
name='Pool cross-correlation')
if unary_control:
logger.info("Only one control supplied.")
if not simplicate_experiment:
logger.info("Using one control for both replicate 1 and 2 and for the pool.")
rep2_control = rep1_control
control_for_pool = rep1_control
pool_ctl_msg = "one control"
else:
pool_controls_subjob = pool_applet.run(
{"inputs": [ctl1_ta, ctl2_ta],
"prefix": "PL_ctls"},
name='Pool controls')
pooled_controls = pool_controls_subjob.get_output_ref("pooled")
# always use the pooled controls for the pool
control_for_pool = pooled_controls
pool_ctl_msg = "pooled controls"
# use the pooled controls for the reps depending on the ratio of rep to
# control reads
ratio_ctl_reads = float(ntags_ctl1)/float(ntags_ctl2)
if ratio_ctl_reads < 1:
ratio_ctl_reads = 1/ratio_ctl_reads
ratio_cutoff = 1.2
if ratio_ctl_reads > ratio_cutoff:
logger.info(
"Number of reads in controls differ by > factor of %f. Using pooled controls."
% (ratio_cutoff))
rep1_control = pooled_controls
rep2_control = pooled_controls
else:
if ntags_ctl1 < ntags_rep1:
logger.info("Fewer reads in control replicate 1 than experiment replicate 1. Using pooled controls for replicate 1.")
rep1_control = pooled_controls
rep1_ctl_msg = "pooled controls"
elif not simplicate_experiment and ntags_ctl2 < ntags_rep2:
logger.info("Fewer reads in control replicate 2 than experiment replicate 2. Using pooled controls for replicate 2.")
rep2_control = pooled_controls
rep2_ctl_msg = "pooled controls"
else:
logger.info(
"Using distinct controls for replicate 1 and 2.")
rep1_control = ctl1_ta # default. May be changed later.
rep2_control = ctl2_ta # default. May be changed later.
rep1_ctl_msg = "control rep1"
rep2_ctl_msg = "control rep2"
pseudoreplicator_applet = dxpy.find_one_data_object(
classname='applet',
name='pseudoreplicator',
zero_ok=False,
more_ok=False,
return_handler=True)
rep1_pr_subjob = pseudoreplicator_applet.run({"input_tags": rep1_ta})
if not simplicate_experiment:
rep2_pr_subjob = pseudoreplicator_applet.run({"input_tags": rep2_ta})
pool_pr1_subjob = pool_applet.run(
{"inputs": [rep1_pr_subjob.get_output_ref("pseudoreplicate1"),
rep2_pr_subjob.get_output_ref("pseudoreplicate1")],
"prefix": 'PPR1'})
pool_pr2_subjob = pool_applet.run(
{"inputs": [rep1_pr_subjob.get_output_ref("pseudoreplicate2"),
rep2_pr_subjob.get_output_ref("pseudoreplicate2")],
"prefix": 'PPR2'})
common_args = {
'chrom_sizes': chrom_sizes,
'genomesize': genomesize,
'narrowpeak_as': narrowpeak_as,
'gappedpeak_as': gappedpeak_as,
'broadpeak_as': broadpeak_as
}
# if the fragment_length argument is given, update macs2 input
if fragment_length is not None:
common_args.update({'fragment_length' : fragment_length})
common_args.update({'prefix': 'r1'})
rep1_peaks_subjob = macs2( rep1_ta,
rep1_control,
rep1_xcor, **common_args)
common_args.update({'prefix': 'r1pr1'})
rep1pr1_peaks_subjob = macs2( rep1_pr_subjob.get_output_ref("pseudoreplicate1"),
rep1_control,
rep1_xcor, **common_args)
common_args.update({'prefix': 'r1pr2'})
rep1pr2_peaks_subjob = macs2( rep1_pr_subjob.get_output_ref("pseudoreplicate2"),
rep1_control,
rep1_xcor, **common_args)
if not simplicate_experiment:
common_args.update({'prefix': 'r2'})
rep2_peaks_subjob = macs2( rep2_ta,
rep2_control,
rep2_xcor, **common_args)
common_args.update({'prefix': 'r2pr1'})
rep2pr1_peaks_subjob = macs2( rep2_pr_subjob.get_output_ref("pseudoreplicate1"),
rep2_control,
rep2_xcor, **common_args)
common_args.update({'prefix': 'r2pr2'})
rep2pr2_peaks_subjob = macs2( rep2_pr_subjob.get_output_ref("pseudoreplicate2"),
rep2_control,
rep2_xcor, **common_args)
common_args.update({'prefix': 'pool'})
pooled_peaks_subjob = macs2( pooled_replicates,
control_for_pool,
pooled_replicates_xcor_subjob.get_output_ref("CC_scores_file"), **common_args)
common_args.update({'prefix': 'ppr1'})
pooledpr1_peaks_subjob = macs2( pool_pr1_subjob.get_output_ref("pooled"),
control_for_pool,
pooled_replicates_xcor_subjob.get_output_ref("CC_scores_file"), **common_args)
common_args.update({'prefix': 'ppr2'})
pooledpr2_peaks_subjob = macs2( pool_pr2_subjob.get_output_ref("pooled"),
control_for_pool,
pooled_replicates_xcor_subjob.get_output_ref("CC_scores_file"), **common_args)
output = {
'rep1_narrowpeaks': rep1_peaks_subjob.get_output_ref("narrowpeaks"),
'rep1_gappedpeaks': rep1_peaks_subjob.get_output_ref("gappedpeaks"),
'rep1_broadpeaks': rep1_peaks_subjob.get_output_ref("broadpeaks"),
'rep1_narrowpeaks_bb': rep1_peaks_subjob.get_output_ref("narrowpeaks_bb"),
'rep1_gappedpeaks_bb': rep1_peaks_subjob.get_output_ref("gappedpeaks_bb"),
'rep1_broadpeaks_bb': rep1_peaks_subjob.get_output_ref("broadpeaks_bb"),
'rep1_fc_signal': rep1_peaks_subjob.get_output_ref("fc_signal"),
'rep1_pvalue_signal': rep1_peaks_subjob.get_output_ref("pvalue_signal"),
'rep1pr1_narrowpeaks': rep1pr1_peaks_subjob.get_output_ref("narrowpeaks"),
'rep1pr1_gappedpeaks': rep1pr1_peaks_subjob.get_output_ref("gappedpeaks"),
'rep1pr1_broadpeaks': rep1pr1_peaks_subjob.get_output_ref("broadpeaks"),
'rep1pr1_fc_signal': rep1pr1_peaks_subjob.get_output_ref("fc_signal"),
'rep1pr1_pvalue_signal': rep1pr1_peaks_subjob.get_output_ref("pvalue_signal"),
'rep1pr2_narrowpeaks': rep1pr2_peaks_subjob.get_output_ref("narrowpeaks"),
'rep1pr2_gappedpeaks': rep1pr2_peaks_subjob.get_output_ref("gappedpeaks"),
'rep1pr2_broadpeaks': rep1pr2_peaks_subjob.get_output_ref("broadpeaks"),
'rep1pr2_fc_signal': rep1pr2_peaks_subjob.get_output_ref("fc_signal"),
'rep1pr2_pvalue_signal': rep1pr2_peaks_subjob.get_output_ref("pvalue_signal")
}
if not simplicate_experiment:
output.update({
'rep2_narrowpeaks': rep2_peaks_subjob.get_output_ref("narrowpeaks"),
'rep2_gappedpeaks': rep2_peaks_subjob.get_output_ref("gappedpeaks"),
'rep2_broadpeaks': rep2_peaks_subjob.get_output_ref("broadpeaks"),
'rep2_narrowpeaks_bb': rep2_peaks_subjob.get_output_ref("narrowpeaks_bb"),
'rep2_gappedpeaks_bb': rep2_peaks_subjob.get_output_ref("gappedpeaks_bb"),
'rep2_broadpeaks_bb': rep2_peaks_subjob.get_output_ref("broadpeaks_bb"),
'rep2_fc_signal': rep2_peaks_subjob.get_output_ref("fc_signal"),
'rep2_pvalue_signal': rep2_peaks_subjob.get_output_ref("pvalue_signal"),
'rep2pr1_narrowpeaks': rep2pr1_peaks_subjob.get_output_ref("narrowpeaks"),
'rep2pr1_gappedpeaks': rep2pr1_peaks_subjob.get_output_ref("gappedpeaks"),
'rep2pr1_broadpeaks': rep2pr1_peaks_subjob.get_output_ref("broadpeaks"),
'rep2pr1_fc_signal': rep2pr1_peaks_subjob.get_output_ref("fc_signal"),
'rep2pr1_pvalue_signal': rep2pr1_peaks_subjob.get_output_ref("pvalue_signal"),
'rep2pr2_narrowpeaks': rep2pr2_peaks_subjob.get_output_ref("narrowpeaks"),
'rep2pr2_gappedpeaks': rep2pr2_peaks_subjob.get_output_ref("gappedpeaks"),
'rep2pr2_broadpeaks': rep2pr2_peaks_subjob.get_output_ref("broadpeaks"),
'rep2pr2_fc_signal': rep2pr2_peaks_subjob.get_output_ref("fc_signal"),
'rep2pr2_pvalue_signal': rep2pr2_peaks_subjob.get_output_ref("pvalue_signal"),
'pooled_narrowpeaks': pooled_peaks_subjob.get_output_ref("narrowpeaks"),
'pooled_gappedpeaks': pooled_peaks_subjob.get_output_ref("gappedpeaks"),
'pooled_broadpeaks': pooled_peaks_subjob.get_output_ref("broadpeaks"),
'pooled_narrowpeaks_bb': pooled_peaks_subjob.get_output_ref("narrowpeaks_bb"),
'pooled_gappedpeaks_bb': pooled_peaks_subjob.get_output_ref("gappedpeaks_bb"),
'pooled_broadpeaks_bb': pooled_peaks_subjob.get_output_ref("broadpeaks_bb"),
'pooled_fc_signal': pooled_peaks_subjob.get_output_ref("fc_signal"),
'pooled_pvalue_signal': pooled_peaks_subjob.get_output_ref("pvalue_signal"),
'pooledpr1_narrowpeaks': pooledpr1_peaks_subjob.get_output_ref("narrowpeaks"),
'pooledpr1_gappedpeaks': pooledpr1_peaks_subjob.get_output_ref("gappedpeaks"),
'pooledpr1_broadpeaks': pooledpr1_peaks_subjob.get_output_ref("broadpeaks"),
'pooledpr1_fc_signal': pooledpr1_peaks_subjob.get_output_ref("fc_signal"),
'pooledpr1_pvalue_signal': pooledpr1_peaks_subjob.get_output_ref("pvalue_signal"),
'pooledpr2_narrowpeaks': pooledpr2_peaks_subjob.get_output_ref("narrowpeaks"),
'pooledpr2_gappedpeaks': pooledpr2_peaks_subjob.get_output_ref("gappedpeaks"),
'pooledpr2_broadpeaks': pooledpr2_peaks_subjob.get_output_ref("broadpeaks"),
'pooledpr2_fc_signal': pooledpr2_peaks_subjob.get_output_ref("fc_signal"),
'pooledpr2_pvalue_signal': pooledpr2_peaks_subjob.get_output_ref("pvalue_signal")
})
return output
def main(rep1_ta,
rep2_ta,
ctl1_ta,
ctl2_ta,
rep1_xcor,
rep2_xcor,
npeaks,
nodups,
rep1_paired_end,
rep2_paired_end,
chrom_sizes,
as_file=None,
idr_peaks=False):
if not rep1_paired_end == rep2_paired_end:
raise ValueError('Mixed PE/SE not supported (yet)')
paired_end = rep1_paired_end
rep1_ta_file = dxpy.DXFile(rep1_ta)
rep2_ta_file = dxpy.DXFile(rep2_ta)
unary_control = ctl1_ta == ctl2_ta
ctl1_ta_file = dxpy.DXFile(ctl1_ta)
ctl2_ta_file = dxpy.DXFile(ctl2_ta)
rep1_xcor_file = dxpy.DXFile(rep1_xcor)
rep2_xcor_file = dxpy.DXFile(rep2_xcor)
dxpy.download_dxfile(rep1_ta_file.get_id(), rep1_ta_file.name)
dxpy.download_dxfile(rep2_ta_file.get_id(), rep2_ta_file.name)
dxpy.download_dxfile(ctl1_ta_file.get_id(), ctl1_ta_file.name)
dxpy.download_dxfile(ctl2_ta_file.get_id(), ctl2_ta_file.name)
dxpy.download_dxfile(rep1_xcor_file.get_id(), rep1_xcor_file.name)
dxpy.download_dxfile(rep2_xcor_file.get_id(), rep2_xcor_file.name)
rep1_ta_filename = rep1_ta_file.name
rep2_ta_filename = rep2_ta_file.name
ctl1_ta_filename = ctl1_ta_file.name
ctl2_ta_filename = ctl2_ta_file.name
ntags_rep1 = common.count_lines(rep1_ta_filename)
ntags_rep2 = common.count_lines(rep2_ta_filename)
ntags_ctl1 = common.count_lines(ctl1_ta_filename)
ntags_ctl2 = common.count_lines(ctl2_ta_filename)
for n, name, filename in [(ntags_rep1, 'replicate 1', rep1_ta_filename),
(ntags_rep2, 'replicate 2', rep2_ta_filename),
(ntags_ctl1, 'control 1', ctl1_ta_filename),
(ntags_ctl2, 'control 2', ctl2_ta_filename)]:
print("Found %d tags in %s file %s" % (n, name, filename))
print(
subprocess.check_output('ls -l', shell=True, stderr=subprocess.STDOUT))
pool_applet = dxpy.find_one_data_object(classname='applet',
name='pool',
project=dxpy.PROJECT_CONTEXT_ID,
zero_ok=False,
more_ok=False,
return_handler=True)
pool_replicates_subjob = \
pool_applet.run(
{"inputs": [rep1_ta, rep2_ta]},
name='Pool replicates')
pooled_replicates = pool_replicates_subjob.get_output_ref("pooled")
pooled_replicates_xcor_subjob = \
xcor_only(
pooled_replicates,
paired_end,
name='Pool cross-correlation')
rep1_control = ctl1_ta # default. May be changed later.
rep1_ctl_msg = "control rep1"
rep2_control = ctl2_ta # default. May be changed later.
rep2_ctl_msg = "control rep2"
if unary_control:
print(
"Only one control supplied. Using it for both replicate 1 and 2 and for the pool."
)
control_for_pool = rep1_control
pool_ctl_msg = "one control"
else:
pool_controls_subjob = \
pool_applet.run(
{"inputs": [ctl1_ta, ctl2_ta]},
name='Pool controls')
pooled_controls = pool_controls_subjob.get_output_ref("pooled")
# always use the pooled controls for the pool
control_for_pool = pooled_controls
pool_ctl_msg = "pooled controls"
# use the pooled controls for the reps depending on the ratio of
# rep to control reads
ratio_ctl_reads = float(ntags_ctl1) / float(ntags_ctl2)
if ratio_ctl_reads < 1:
ratio_ctl_reads = 1 / ratio_ctl_reads
ratio_cutoff = 1.2
if ratio_ctl_reads > ratio_cutoff:
print(
"Number of reads in controls differ by > factor of %f. Using pooled controls."
% (ratio_cutoff))
rep1_control = pooled_controls
rep1_ctl_msg = "pooled controls"
rep2_control = pooled_controls
rep2_ctl_msg = "pooled controls"
else:
if ntags_ctl1 < ntags_rep1:
print(
"Fewer reads in control replicate 1 than experiment replicate 1. Using pooled controls for replicate 1."
)
rep1_control = pooled_controls
rep1_ctl_msg = "pooled controls"
elif ntags_ctl2 < ntags_rep2:
print(
"Fewer reads in control replicate 2 than experiment replicate 2. Using pooled controls for replicate 2."
)
rep2_control = pooled_controls
rep2_ctl_msg = "pooled controls"
else:
print("Using distinct controls for replicate 1 and 2.")
rep1_ctl_msg = "control rep1"
rep2_ctl_msg = "control rep2"
rep1_peaks_subjob = spp(rep1_ta,
rep1_control,
rep1_xcor,
chrom_sizes=chrom_sizes,
bigbed=True,
as_file=as_file,
name='Rep1 peaks vs %s' % (rep1_ctl_msg))
rep2_peaks_subjob = spp(rep2_ta,
rep2_control,
rep2_xcor,
chrom_sizes=chrom_sizes,
bigbed=True,
as_file=as_file,
name='Rep2 peaks vs %s' % (rep2_ctl_msg))
pooled_peaks_subjob = spp(
pooled_replicates,
control_for_pool,
pooled_replicates_xcor_subjob.get_output_ref("CC_scores_file"),
chrom_sizes=chrom_sizes,
bigbed=True,
as_file=as_file,
name='Pooled peaks vs %s' % (pool_ctl_msg))
output = {
'rep1_peaks': rep1_peaks_subjob.get_output_ref("peaks"),
'rep1_peaks_bb': rep1_peaks_subjob.get_output_ref("peaks_bb"),
'rep1_xcor_plot': rep1_peaks_subjob.get_output_ref("xcor_plot"),
'rep1_xcor_scores': rep1_peaks_subjob.get_output_ref("xcor_scores"),
'rep2_peaks': rep2_peaks_subjob.get_output_ref("peaks"),
'rep2_peaks_bb': rep2_peaks_subjob.get_output_ref("peaks_bb"),
'rep2_xcor_plot': rep2_peaks_subjob.get_output_ref("xcor_plot"),
'rep2_xcor_scores': rep2_peaks_subjob.get_output_ref("xcor_scores"),
'pooled_peaks': pooled_peaks_subjob.get_output_ref("peaks"),
'pooled_peaks_bb': pooled_peaks_subjob.get_output_ref("peaks_bb"),
'pooled_xcor_plot': pooled_peaks_subjob.get_output_ref("xcor_plot"),
'pooled_xcor_scores': pooled_peaks_subjob.get_output_ref("xcor_scores")
}
if idr_peaks: # also call peaks on pseudoreplicates for IDR
pseudoreplicator_applet = \
dxpy.find_one_data_object(
classname='applet',
name='pseudoreplicator',
project=dxpy.PROJECT_CONTEXT_ID,
zero_ok=False,
more_ok=False,
return_handler=True)
rep1_pr_subjob = \
pseudoreplicator_applet.run(
{"input_tags": rep1_ta},
name='Pseudoreplicate rep1 -> R1PR1,2')
rep2_pr_subjob = \
pseudoreplicator_applet.run(
{"input_tags": rep2_ta},
name='Pseudoreplicate rep2 -> R2PR1,2')
pool_pr1_subjob = pool_applet.run(
{
"inputs": [
rep1_pr_subjob.get_output_ref("pseudoreplicate1"),
rep2_pr_subjob.get_output_ref("pseudoreplicate1")
]
},
name='Pool R1PR1+R2PR1 -> PPR1')
pool_pr2_subjob = pool_applet.run(
{
"inputs": [
rep1_pr_subjob.get_output_ref("pseudoreplicate2"),
rep2_pr_subjob.get_output_ref("pseudoreplicate2")
]
},
name='Pool R1PR2+R2PR2 -> PPR2')
rep1_pr1_xcor_subjob = xcor_only(
rep1_pr_subjob.get_output_ref("pseudoreplicate1"),
paired_end,
name='R1PR1 cross-correlation')
rep1_pr2_xcor_subjob = xcor_only(
rep1_pr_subjob.get_output_ref("pseudoreplicate2"),
paired_end,
name='R1PR2 cross-correlation')
rep2_pr1_xcor_subjob = xcor_only(
rep2_pr_subjob.get_output_ref("pseudoreplicate1"),
paired_end,
name='R2PR1 cross-correlation')
rep2_pr2_xcor_subjob = xcor_only(
rep2_pr_subjob.get_output_ref("pseudoreplicate2"),
paired_end,
name='R2PR2 cross-correlation')
pool_pr1_xcor_subjob = xcor_only(
pool_pr1_subjob.get_output_ref("pooled"),
paired_end,
name='PPR1 cross-correlation')
pool_pr2_xcor_subjob = xcor_only(
pool_pr2_subjob.get_output_ref("pooled"),
paired_end,
name='PPR2 cross-correlation')
rep1pr1_peaks_subjob = spp(
rep1_pr_subjob.get_output_ref("pseudoreplicate1"),
rep1_control,
rep1_pr1_xcor_subjob.get_output_ref("CC_scores_file"),
chrom_sizes=chrom_sizes,
bigbed=False,
name='R1PR1 peaks vs %s' % (rep1_ctl_msg))
rep1pr2_peaks_subjob = spp(
rep1_pr_subjob.get_output_ref("pseudoreplicate2"),
rep1_control,
rep1_pr2_xcor_subjob.get_output_ref("CC_scores_file"),
chrom_sizes=chrom_sizes,
bigbed=False,
name='R1PR2 peaks vs %s' % (rep1_ctl_msg))
rep2pr1_peaks_subjob = spp(
rep2_pr_subjob.get_output_ref("pseudoreplicate1"),
rep2_control,
rep2_pr1_xcor_subjob.get_output_ref("CC_scores_file"),
chrom_sizes=chrom_sizes,
bigbed=False,
name='R2PR1 peaks vs %s' % (rep2_ctl_msg))
rep2pr2_peaks_subjob = spp(
rep2_pr_subjob.get_output_ref("pseudoreplicate2"),
rep2_control,
rep2_pr2_xcor_subjob.get_output_ref("CC_scores_file"),
chrom_sizes=chrom_sizes,
bigbed=False,
name='R2PR2 peaks vs %s' % (rep2_ctl_msg))
pooledpr1_peaks_subjob = spp(
pool_pr1_subjob.get_output_ref("pooled"),
control_for_pool,
pool_pr1_xcor_subjob.get_output_ref("CC_scores_file"),
chrom_sizes=chrom_sizes,
bigbed=False,
name='PPR1 peaks vs %s' % (pool_ctl_msg))
pooledpr2_peaks_subjob = spp(
pool_pr2_subjob.get_output_ref("pooled"),
control_for_pool,
pool_pr2_xcor_subjob.get_output_ref("CC_scores_file"),
chrom_sizes=chrom_sizes,
bigbed=False,
name='PPR2 peaks vs %s' % (pool_ctl_msg))
output.update({
'rep1pr1_peaks':
rep1pr1_peaks_subjob.get_output_ref("peaks"),
'rep1pr1_xcor_plot':
rep1pr1_peaks_subjob.get_output_ref("xcor_plot"),
'rep1pr1_xcor_scores':
rep1pr1_peaks_subjob.get_output_ref("xcor_scores"),
'rep1pr2_peaks':
rep1pr2_peaks_subjob.get_output_ref("peaks"),
'rep1pr2_xcor_plot':
rep1pr2_peaks_subjob.get_output_ref("xcor_plot"),
'rep1pr2_xcor_scores':
rep1pr2_peaks_subjob.get_output_ref("xcor_scores"),
'rep2pr1_peaks':
rep2pr1_peaks_subjob.get_output_ref("peaks"),
'rep2pr1_xcor_plot':
rep2pr1_peaks_subjob.get_output_ref("xcor_plot"),
'rep2pr1_xcor_scores':
rep2pr1_peaks_subjob.get_output_ref("xcor_scores"),
'rep2pr2_peaks':
rep2pr2_peaks_subjob.get_output_ref("peaks"),
'rep2pr2_xcor_plot':
rep2pr2_peaks_subjob.get_output_ref("xcor_plot"),
'rep2pr2_xcor_scores':
rep2pr2_peaks_subjob.get_output_ref("xcor_scores"),
'pooledpr1_peaks':
pooledpr1_peaks_subjob.get_output_ref("peaks"),
'pooledpr1_xcor_plot':
pooledpr1_peaks_subjob.get_output_ref("xcor_plot"),
'pooledpr1_xcor_scores':
pooledpr1_peaks_subjob.get_output_ref("xcor_scores"),
'pooledpr2_peaks':
pooledpr2_peaks_subjob.get_output_ref("peaks"),
'pooledpr2_xcor_plot':
pooledpr2_peaks_subjob.get_output_ref("xcor_plot"),
'pooledpr2_xcor_scores':
pooledpr2_peaks_subjob.get_output_ref("xcor_scores"),
})
return output
def main(experiment,
reps_peaks,
r1pr_peaks,
r2pr_peaks,
pooledpr_peaks,
chrom_sizes,
as_file,
blacklist=None):
#TODO for now just taking the peak files. This applet should actually call IDR instead of
#putting that in the workflow populator script
# Initialize the data object inputs on the platform into
# dxpy.DXDataObject instances.
reps_peaks_file = dxpy.DXFile(reps_peaks)
r1pr_peaks_file = dxpy.DXFile(r1pr_peaks)
r2pr_peaks_file = dxpy.DXFile(r2pr_peaks)
pooledpr_peaks_file = dxpy.DXFile(pooledpr_peaks)
chrom_sizes_file = dxpy.DXFile(chrom_sizes)
as_file_file = dxpy.DXFile(as_file)
if blacklist is not None:
blacklist_file = dxpy.DXFile(blacklist)
blacklist_filename = 'blacklist_%s' % (blacklist_file.name)
dxpy.download_dxfile(blacklist_file.get_id(), blacklist_filename)
blacklist_filename = common.uncompress(blacklist_filename)
# Download the file inputs to the local file system.
#Need to prepend something to ensure the local filenames will be unique
reps_peaks_filename = 'true_%s' % (reps_peaks_file.name)
r1pr_peaks_filename = 'r1pr_%s' % (r1pr_peaks_file.name)
r2pr_peaks_filename = 'r2pr_%s' % (r2pr_peaks_file.name)
pooledpr_peaks_filename = 'pooledpr_%s' % (pooledpr_peaks_file.name)
chrom_sizes_filename = chrom_sizes_file.name
as_file_filename = as_file_file.name
dxpy.download_dxfile(reps_peaks_file.get_id(), reps_peaks_filename)
dxpy.download_dxfile(r1pr_peaks_file.get_id(), r1pr_peaks_filename)
dxpy.download_dxfile(r2pr_peaks_file.get_id(), r2pr_peaks_filename)
dxpy.download_dxfile(pooledpr_peaks_file.get_id(), pooledpr_peaks_filename)
dxpy.download_dxfile(chrom_sizes_file.get_id(), chrom_sizes_filename)
dxpy.download_dxfile(as_file_file.get_id(), as_file_filename)
print subprocess.check_output('ls -l', shell=True)
reps_peaks_filename = common.uncompress(reps_peaks_filename)
r1pr_peaks_filename = common.uncompress(r1pr_peaks_filename)
r2pr_peaks_filename = common.uncompress(r2pr_peaks_filename)
pooledpr_peaks_filename = common.uncompress(pooledpr_peaks_filename)
Nt = common.count_lines(reps_peaks_filename)
print "%d peaks from true replicates" % (Nt)
N1 = common.count_lines(r1pr_peaks_filename)
print "%d peaks from rep1 self-pseudoreplicates" % (N1)
N2 = common.count_lines(r2pr_peaks_filename)
print "%d peaks from rep2 self-pseudoreplicates" % (N2)
Np = common.count_lines(pooledpr_peaks_filename)
print "%d peaks from pooled pseudoreplicates" % (Np)
conservative_set_filename = '%s_final_conservative.narrowPeak' % (
experiment)
if blacklist is not None:
blacklist_filter(reps_peaks_filename, conservative_set_filename,
blacklist_filename)
else:
conservative_set_filename = reps_peaks_filename
Ncb = common.count_lines(conservative_set_filename)
print "%d peaks blacklisted from the conservative set" % (Nt - Ncb)
if Nt >= Np:
peaks_to_filter_filename = reps_peaks_filename
No = Nt
else:
peaks_to_filter_filename = pooledpr_peaks_filename
No = Np
optimal_set_filename = '%s_final_optimal.narrowPeak' % (experiment)
if blacklist is not None:
blacklist_filter(peaks_to_filter_filename, optimal_set_filename,
blacklist_filename)
else:
optimal_set_filename = peaks_to_filter_filename
Nob = common.count_lines(optimal_set_filename)
print "%d peaks blacklisted from the optimal set" % (No - Nob)
rescue_ratio = float(max(Np, Nt)) / float(min(Np, Nt))
self_consistency_ratio = float(max(N1, N2)) / float(min(N1, N2))
if rescue_ratio > 2 and self_consistency_ratio > 2:
reproducibility = 'fail'
elif rescue_ratio > 2 or self_consistency_ratio > 2:
reproducibility = 'borderline'
else:
reproducibility = 'pass'
output = {}
#bedtobigbed often fails, so skip creating the bb if it does
conservative_set_bb_filename = common.bed2bb(conservative_set_filename,
chrom_sizes_filename,
as_file_filename)
optimal_set_bb_filename = common.bed2bb(optimal_set_filename,
chrom_sizes_filename,
as_file_filename)
if conservative_set_bb_filename:
conservative_set_bb_output = dxpy.upload_local_file(
conservative_set_bb_filename)
output.update(
{"conservative_set_bb": dxpy.dxlink(conservative_set_bb_output)})
if optimal_set_bb_filename:
optimal_set_bb_output = dxpy.upload_local_file(optimal_set_bb_filename)
output.update({"optimal_set_bb": dxpy.dxlink(optimal_set_bb_output)})
output.update({
"Nt":
Nt,
"N1":
N1,
"N2":
N2,
"Np":
Np,
"conservative_set":
dxpy.dxlink(
dxpy.upload_local_file(
common.compress(conservative_set_filename))),
"optimal_set":
dxpy.dxlink(
dxpy.upload_local_file(common.compress(optimal_set_filename))),
"rescue_ratio":
rescue_ratio,
"self_consistency_ratio":
self_consistency_ratio,
"reproducibility_test":
reproducibility
})
logging.info("Exiting with output: %s", output)
return output
def replicated_overlap(rep1_peaks, rep2_peaks, pooled_peaks,
pooledpr1_peaks, pooledpr2_peaks,
rep1_ta, rep1_xcor, rep2_ta, rep2_xcor,
paired_end, chrom_sizes, as_file, peak_type, prefix,
fragment_length=None):
rep1_peaks_file = dxpy.DXFile(rep1_peaks)
rep2_peaks_file = dxpy.DXFile(rep2_peaks)
pooled_peaks_file = dxpy.DXFile(pooled_peaks)
pooledpr1_peaks_file = dxpy.DXFile(pooledpr1_peaks)
pooledpr2_peaks_file = dxpy.DXFile(pooledpr2_peaks)
rep1_ta_file = dxpy.DXFile(rep1_ta)
rep2_ta_file = dxpy.DXFile(rep2_ta)
rep1_xcor_file = dxpy.DXFile(rep1_xcor)
rep2_xcor_file = dxpy.DXFile(rep2_xcor)
chrom_sizes_file = dxpy.DXFile(chrom_sizes)
as_file_file = dxpy.DXFile(as_file)
# Input filenames - necessary to define each explicitly because input files
# could have the same name, in which case subsequent
# file would overwrite previous file
rep1_peaks_fn = 'rep1-%s' % (rep1_peaks_file.name)
rep2_peaks_fn = 'rep2-%s' % (rep2_peaks_file.name)
pooled_peaks_fn = 'pooled-%s' % (pooled_peaks_file.name)
pooledpr1_peaks_fn = 'pooledpr1-%s' % (pooledpr1_peaks_file.name)
pooledpr2_peaks_fn = 'pooledpr2-%s' % (pooledpr2_peaks_file.name)
rep1_ta_fn = 'r1ta_%s' % (rep1_ta_file.name)
rep2_ta_fn = 'r2ta_%s' % (rep2_ta_file.name)
rep1_xcor_fn = 'r1cc_%s' % (rep1_xcor_file.name)
rep2_xcor_fn = 'r2cc_%s' % (rep2_xcor_file.name)
chrom_sizes_fn = 'chrom.sizes'
as_file_fn = '%s.as' % (peak_type)
# Output filenames
if prefix:
basename = prefix
else:
# strip off the peak and compression extensions
m = re.match(
'(.*)(\.%s)+(\.((gz)|(Z)|(bz)|(bz2)))' % (peak_type),
pooled_peaks.name)
if m:
basename = m.group(1)
else:
basename = pooled_peaks.name
overlapping_peaks_fn = '%s.replicated.%s' % (basename, peak_type)
overlapping_peaks_bb_fn = overlapping_peaks_fn + '.bb'
rejected_peaks_fn = '%s.rejected.%s' % (basename, peak_type)
rejected_peaks_bb_fn = rejected_peaks_fn + '.bb'
# Intermediate filenames
overlap_tr_fn = 'replicated_tr.%s' % (peak_type)
overlap_pr_fn = 'replicated_pr.%s' % (peak_type)
# Download file inputs to the local file system with local filenames
dxpy.download_dxfile(rep1_peaks_file.get_id(), rep1_peaks_fn)
dxpy.download_dxfile(rep2_peaks_file.get_id(), rep2_peaks_fn)
dxpy.download_dxfile(pooled_peaks_file.get_id(), pooled_peaks_fn)
dxpy.download_dxfile(pooledpr1_peaks_file.get_id(), pooledpr1_peaks_fn)
dxpy.download_dxfile(pooledpr2_peaks_file.get_id(), pooledpr2_peaks_fn)
dxpy.download_dxfile(rep1_ta_file.get_id(), rep1_ta_fn)
dxpy.download_dxfile(rep2_ta_file.get_id(), rep2_ta_fn)
dxpy.download_dxfile(rep1_xcor_file.get_id(), rep1_xcor_fn)
dxpy.download_dxfile(rep2_xcor_file.get_id(), rep2_xcor_fn)
dxpy.download_dxfile(chrom_sizes_file.get_id(), chrom_sizes_fn)
dxpy.download_dxfile(as_file_file.get_id(), as_file_fn)
pool_applet = dxpy.find_one_data_object(
classname='applet',
name='pool',
project=dxpy.PROJECT_CONTEXT_ID,
zero_ok=False,
more_ok=False,
return_handler=True)
pool_replicates_subjob = \
pool_applet.run(
{"inputs": [rep1_ta, rep2_ta],
"prefix": 'pooled_reps'},
name='Pool replicates')
# If fragment length was given by user we skip pooled_replicates
# _xcor_subjob, set the pool_xcor_filename to None, and update
# the flag fragment_length_given_by_user. Otherwise, run the subjob
# to be able to extract the fragment length fron cross-correlations.
if fragment_length is not None:
pool_xcor_filename = None
fraglen = fragment_length
fragment_length_given_by_user = True
else:
pooled_replicates_xcor_subjob = \
xcor_only(
pool_replicates_subjob.get_output_ref("pooled"),
paired_end,
spp_version=None,
name='Pool cross-correlation')
pooled_replicates_xcor_subjob.wait_on_done()
pool_xcor_link = pooled_replicates_xcor_subjob.describe()['output'].get("CC_scores_file")
pool_xcor_file = dxpy.get_handler(pool_xcor_link)
pool_xcor_filename = 'poolcc_%s' % (pool_xcor_file.name)
dxpy.download_dxfile(pool_xcor_file.get_id(), pool_xcor_filename)
fraglen = common.xcor_fraglen(pool_xcor_filename)
fragment_length_given_by_user = False
pool_replicates_subjob.wait_on_done()
pool_ta_link = pool_replicates_subjob.describe()['output'].get("pooled")
pool_ta_file = dxpy.get_handler(pool_ta_link)
pool_ta_filename = 'poolta_%s' % (pool_ta_file.name)
dxpy.download_dxfile(pool_ta_file.get_id(), pool_ta_filename)
logger.info(subprocess.check_output('set -x; ls -l', shell=True))
# the only difference between the peak_types is how the extra columns are
# handled
if peak_type == "narrowPeak":
awk_command = r"""awk 'BEGIN{FS="\t";OFS="\t"}{s1=$3-$2; s2=$13-$12; if (($21/s1 >= 0.5) || ($21/s2 >= 0.5)) {print $0}}'"""
cut_command = 'cut -f 1-10'
bed_type = 'bed6+4'
elif peak_type == "gappedPeak":
awk_command = r"""awk 'BEGIN{FS="\t";OFS="\t"}{s1=$3-$2; s2=$18-$17; if (($31/s1 >= 0.5) || ($31/s2 >= 0.5)) {print $0}}'"""
cut_command = 'cut -f 1-15'
bed_type = 'bed12+3'
elif peak_type == "broadPeak":
awk_command = r"""awk 'BEGIN{FS="\t";OFS="\t"}{s1=$3-$2; s2=$12-$11; if (($19/s1 >= 0.5) || ($19/s2 >= 0.5)) {print $0}}'"""
cut_command = 'cut -f 1-9'
bed_type = 'bed6+3'
else:
assert peak_type in ['narrowPeak', 'gappedPeak', 'broadPeak'], "%s is unrecognized. peak_type should be narrowPeak, gappedPeak or broadPeak." % (peak_type)
# Find pooled peaks that overlap Rep1 and Rep2 where overlap is defined as
# the fractional overlap wrt any one of the overlapping peak pairs > 0.5
out, err = common.run_pipe([
'intersectBed -wo -a %s -b %s' % (pooled_peaks_fn, rep1_peaks_fn),
awk_command,
cut_command,
'sort -u',
'intersectBed -wo -a stdin -b %s' % (rep2_peaks_fn),
awk_command,
cut_command,
'sort -u'
], overlap_tr_fn)
print(
"%d peaks overlap with both true replicates"
% (common.count_lines(overlap_tr_fn)))
# Find pooled peaks that overlap PseudoRep1 and PseudoRep2 where
# overlap is defined as the fractional overlap wrt any one of the
# overlapping peak pairs > 0.5
out, err = common.run_pipe([
'intersectBed -wo -a %s -b %s' % (pooled_peaks_fn, pooledpr1_peaks_fn),
awk_command,
cut_command,
'sort -u',
'intersectBed -wo -a stdin -b %s' % (pooledpr2_peaks_fn),
awk_command,
cut_command,
'sort -u'
], overlap_pr_fn)
print(
"%d peaks overlap with both pooled pseudoreplicates"
% (common.count_lines(overlap_pr_fn)))
# Combine peak lists
out, err = common.run_pipe([
'cat %s %s' % (overlap_tr_fn, overlap_pr_fn),
'sort -u'
], overlapping_peaks_fn)
print(
"%d peaks overlap with true replicates or with pooled pseudoreplicates"
% (common.count_lines(overlapping_peaks_fn)))
# rejected peaks
out, err = common.run_pipe([
'intersectBed -wa -v -a %s -b %s' % (pooled_peaks_fn, overlapping_peaks_fn)
], rejected_peaks_fn)
print("%d peaks were rejected" % (common.count_lines(rejected_peaks_fn)))
# calculate FRiP (Fraction of Reads in Peaks)
reads_in_peaks_fn = 'reads_in_%s.ta' % (peak_type)
n_reads, n_reads_in_peaks, frip_score = common.frip(
pool_ta_filename, pool_xcor_filename, overlapping_peaks_fn,
chrom_sizes_fn, fraglen, reads_in_peaks_fn=reads_in_peaks_fn)
# count peaks
npeaks_in = common.count_lines(common.uncompress(pooled_peaks_fn))
npeaks_out = common.count_lines(overlapping_peaks_fn)
npeaks_rejected = common.count_lines(rejected_peaks_fn)
# make bigBed files for visualization
overlapping_peaks_bb_fn = common.bed2bb(
overlapping_peaks_fn, chrom_sizes_fn, as_file_fn, bed_type=bed_type)
rejected_peaks_bb_fn = common.bed2bb(
rejected_peaks_fn, chrom_sizes_fn, as_file_fn, bed_type=bed_type)
# Upload file outputs from the local file system.
overlapping_peaks = dxpy.upload_local_file(common.compress(overlapping_peaks_fn))
overlapping_peaks_bb = dxpy.upload_local_file(overlapping_peaks_bb_fn)
rejected_peaks = dxpy.upload_local_file(common.compress(rejected_peaks_fn))
rejected_peaks_bb = dxpy.upload_local_file(rejected_peaks_bb_fn)
output = {
"overlapping_peaks" : dxpy.dxlink(overlapping_peaks),
"overlapping_peaks_bb" : dxpy.dxlink(overlapping_peaks_bb),
"rejected_peaks" : dxpy.dxlink(rejected_peaks),
"rejected_peaks_bb" : dxpy.dxlink(rejected_peaks_bb),
"npeaks_in" : npeaks_in,
"npeaks_out" : npeaks_out,
"npeaks_rejected" : npeaks_rejected,
"frip_nreads" : n_reads,
"frip_nreads_in_peaks" : n_reads_in_peaks,
"frip_score" : frip_score,
"fragment_length_used" : fraglen,
"fragment_length_given_by_user": fragment_length_given_by_user
}
return output
def internal_pseudoreplicate_overlap(rep1_peaks, rep2_peaks, pooled_peaks,
rep1_ta, rep1_xcor,
paired_end, chrom_sizes, as_file,
peak_type, prefix, fragment_length=None):
rep1_peaks_file = dxpy.DXFile(rep1_peaks)
rep2_peaks_file = dxpy.DXFile(rep2_peaks)
pooled_peaks_file = dxpy.DXFile(pooled_peaks)
rep1_ta_file = dxpy.DXFile(rep1_ta)
rep1_xcor_file = dxpy.DXFile(rep1_xcor)
chrom_sizes_file = dxpy.DXFile(chrom_sizes)
as_file_file = dxpy.DXFile(as_file)
# Input filenames - necessary to define each explicitly because input files
# could have the same name, in which case subsequent
# file would overwrite previous file
rep1_peaks_fn = 'rep1-%s' % (rep1_peaks_file.name)
rep2_peaks_fn = 'rep2-%s' % (rep2_peaks_file.name)
pooled_peaks_fn = 'pooled-%s' % (pooled_peaks_file.name)
rep1_ta_fn = 'r1ta_%s' % (rep1_ta_file.name)
rep1_xcor_fn = 'r1xc_%s' % (rep1_xcor_file.name)
chrom_sizes_fn = 'chrom.sizes'
as_file_fn = '%s.as' % (peak_type)
# Output filenames
if prefix:
basename = prefix
else:
# strip off the peak and compression extensions
m = re.match(
'(.*)(\.%s)+(\.((gz)|(Z)|(bz)|(bz2)))' % (peak_type),
pooled_peaks.name)
if m:
basename = m.group(1)
else:
basename = pooled_peaks.name
overlapping_peaks_fn = '%s.replicated.%s' % (basename, peak_type)
overlapping_peaks_bb_fn = overlapping_peaks_fn + '.bb'
rejected_peaks_fn = '%s.rejected.%s' % (basename, peak_type)
rejected_peaks_bb_fn = rejected_peaks_fn + '.bb'
# Intermediate filenames
overlap_tr_fn = 'replicated_tr.%s' % (peak_type)
overlap_pr_fn = 'replicated_pr.%s' % (peak_type)
# Download file inputs to the local file system with local filenames
dxpy.download_dxfile(rep1_peaks_file.get_id(), rep1_peaks_fn)
dxpy.download_dxfile(rep2_peaks_file.get_id(), rep2_peaks_fn)
dxpy.download_dxfile(pooled_peaks_file.get_id(), pooled_peaks_fn)
dxpy.download_dxfile(rep1_ta_file.get_id(), rep1_ta_fn)
dxpy.download_dxfile(rep1_xcor_file.get_id(), rep1_xcor_fn)
dxpy.download_dxfile(chrom_sizes_file.get_id(), chrom_sizes_fn)
dxpy.download_dxfile(as_file_file.get_id(), as_file_fn)
logger.info(subprocess.check_output('set -x; ls -l', shell=True))
# the only difference between the peak_types is how the extra columns are
# handled
if peak_type == "narrowPeak":
awk_command = r"""awk 'BEGIN{FS="\t";OFS="\t"}{s1=$3-$2; s2=$13-$12; if (($21/s1 >= 0.5) || ($21/s2 >= 0.5)) {print $0}}'"""
cut_command = 'cut -f 1-10'
bed_type = 'bed6+4'
elif peak_type == "gappedPeak":
awk_command = r"""awk 'BEGIN{FS="\t";OFS="\t"}{s1=$3-$2; s2=$18-$17; if (($31/s1 >= 0.5) || ($31/s2 >= 0.5)) {print $0}}'"""
cut_command = 'cut -f 1-15'
bed_type = 'bed12+3'
elif peak_type == "broadPeak":
awk_command = r"""awk 'BEGIN{FS="\t";OFS="\t"}{s1=$3-$2; s2=$12-$11; if (($19/s1 >= 0.5) || ($19/s2 >= 0.5)) {print $0}}'"""
cut_command = 'cut -f 1-9'
bed_type = 'bed6+3'
else:
assert peak_type in ['narrowPeak', 'gappedPeak', 'broadPeak'], "%s is unrecognized. peak_type should be narrowPeak, gappedPeak or broadPeak." % (peak_type)
# Find pooled peaks that overlap Rep1 and Rep2 where overlap is defined as
# the fractional overlap wrt any one of the overlapping peak pairs > 0.5
out, err = common.run_pipe([
'intersectBed -wo -a %s -b %s' % (pooled_peaks_fn, rep1_peaks_fn),
awk_command,
cut_command,
'sort -u',
'intersectBed -wo -a stdin -b %s' % (rep2_peaks_fn),
awk_command,
cut_command,
'sort -u'
], overlap_tr_fn)
print(
"%d peaks overlap with both true replicates"
% (common.count_lines(overlap_tr_fn)))
# this is a simplicate analysis
# overlapping peaks are just based on pseudoreps of the one pool
out, err = common.run_pipe([
'cat %s' % (overlap_tr_fn),
'sort -u'
], overlapping_peaks_fn)
print(
"%d peaks overlap"
% (common.count_lines(overlapping_peaks_fn)))
# rejected peaks
out, err = common.run_pipe([
'intersectBed -wa -v -a %s -b %s' % (pooled_peaks_fn, overlapping_peaks_fn)
], rejected_peaks_fn)
print("%d peaks were rejected" % (common.count_lines(rejected_peaks_fn)))
# calculate FRiP (Fraction of Reads in Peaks)
# Extract the fragment length estimate from column 3 of the
# cross-correlation scores file or use the user-defined
# fragment_length if given.
if fragment_length is not None:
fraglen = fragment_length
fragment_length_given_by_user = True
else:
fraglen = common.xcor_fraglen(rep1_xcor_fn)
fragment_length_given_by_user = False
# FRiP
reads_in_peaks_fn = 'reads_in_%s.ta' % (peak_type)
n_reads, n_reads_in_peaks, frip_score = common.frip(
rep1_ta_fn, rep1_xcor_fn, overlapping_peaks_fn,
chrom_sizes_fn, fraglen,
reads_in_peaks_fn=reads_in_peaks_fn)
# count peaks
npeaks_in = common.count_lines(common.uncompress(pooled_peaks_fn))
npeaks_out = common.count_lines(overlapping_peaks_fn)
npeaks_rejected = common.count_lines(rejected_peaks_fn)
# make bigBed files for visualization
overlapping_peaks_bb_fn = common.bed2bb(
overlapping_peaks_fn, chrom_sizes_fn, as_file_fn, bed_type=bed_type)
rejected_peaks_bb_fn = common.bed2bb(
rejected_peaks_fn, chrom_sizes_fn, as_file_fn, bed_type=bed_type)
# Upload file outputs from the local file system.
overlapping_peaks = dxpy.upload_local_file(common.compress(overlapping_peaks_fn))
overlapping_peaks_bb = dxpy.upload_local_file(overlapping_peaks_bb_fn)
rejected_peaks = dxpy.upload_local_file(common.compress(rejected_peaks_fn))
rejected_peaks_bb = dxpy.upload_local_file(rejected_peaks_bb_fn)
output = {
"overlapping_peaks" : dxpy.dxlink(overlapping_peaks),
"overlapping_peaks_bb" : dxpy.dxlink(overlapping_peaks_bb),
"rejected_peaks" : dxpy.dxlink(rejected_peaks),
"rejected_peaks_bb" : dxpy.dxlink(rejected_peaks_bb),
"npeaks_in" : npeaks_in,
"npeaks_out" : npeaks_out,
"npeaks_rejected" : npeaks_rejected,
"frip_nreads" : n_reads,
"frip_nreads_in_peaks" : n_reads_in_peaks,
"frip_score" : frip_score,
"fragment_length_used" : fraglen,
"fragment_length_given_by_user": fragment_length_given_by_user
}
return output
def main(rep1_ta,
ctl1_ta,
rep1_xcor,
rep1_paired_end,
chrom_sizes,
genomesize,
narrowpeak_as,
gappedpeak_as,
broadpeak_as,
rep2_ta=None,
ctl2_ta=None,
rep2_xcor=None,
rep2_paired_end=None,
fragment_length=None):
rep1_ta_file = dxpy.DXFile(rep1_ta)
dxpy.download_dxfile(rep1_ta_file.get_id(), rep1_ta_file.name)
rep1_ta_filename = rep1_ta_file.name
ntags_rep1 = common.count_lines(rep1_ta_filename)
simplicate_experiment = rep1_ta and not rep2_ta
if simplicate_experiment:
logger.info(
"No rep2 tags specified so processing as a simplicate experiment.")
else:
logger.info(
"Rep1 and rep2 tags specified so processing as a replicated experiment."
)
if not simplicate_experiment:
assert rep1_paired_end == rep2_paired_end, 'Mixed PE/SE not supported'
rep2_ta_file = dxpy.DXFile(rep2_ta)
dxpy.download_dxfile(rep2_ta_file.get_id(), rep2_ta_file.name)
rep2_ta_filename = rep2_ta_file.name
ntags_rep2 = common.count_lines(rep2_ta_filename)
paired_end = rep1_paired_end
unary_control = (ctl1_ta == ctl2_ta) or not ctl2_ta
ctl1_ta_file = dxpy.DXFile(ctl1_ta)
dxpy.download_dxfile(ctl1_ta_file.get_id(), ctl1_ta_file.name)
ctl1_ta_filename = ctl1_ta_file.name
if not unary_control:
ctl2_ta_file = dxpy.DXFile(ctl2_ta)
dxpy.download_dxfile(ctl2_ta_file.get_id(), ctl2_ta_file.name)
ctl2_ta_filename = ctl2_ta_file.name
else:
ctl2_ta_file = ctl1_ta_file
ctl2_ta_filename = ctl1_ta_file.name
ntags_ctl1 = common.count_lines(ctl1_ta_filename)
ntags_ctl2 = common.count_lines(ctl2_ta_filename)
rep1_control = ctl1_ta # default. May be changed later.
rep1_ctl_msg = "control rep1"
rep2_control = ctl2_ta # default. May be changed later.
rep2_ctl_msg = "control rep2"
rep_info = [(ntags_rep1, 'replicate 1', rep1_ta_filename)]
if not simplicate_experiment:
rep_info.append((ntags_rep2, 'replicate 2', rep2_ta_filename))
rep_info.extend([(ntags_ctl1, 'control 1', ctl1_ta_filename),
(ntags_ctl2, 'control 2', ctl2_ta_filename)])
for n, name, filename in rep_info:
logger.info("Found %d tags in %s file %s" % (n, name, filename))
subprocess.check_output('ls -l', shell=True, stderr=subprocess.STDOUT)
if not simplicate_experiment:
pool_applet = dxpy.find_one_data_object(
classname='applet',
name='pool',
project=dxpy.PROJECT_CONTEXT_ID,
zero_ok=False,
more_ok=False,
return_handler=True)
pool_replicates_subjob = \
pool_applet.run(
{"inputs": [rep1_ta, rep2_ta],
"prefix": 'pooled_reps'},
name='Pool replicates')
pooled_replicates = pool_replicates_subjob.get_output_ref("pooled")
pooled_replicates_xcor_subjob = \
xcor_only(
pooled_replicates,
paired_end,
name='Pool cross-correlation')
if unary_control:
logger.info("Only one control supplied.")
if not simplicate_experiment:
logger.info(
"Using one control for both replicate 1 and 2 and for the pool."
)
rep2_control = rep1_control
control_for_pool = rep1_control
pool_ctl_msg = "one control"
else:
pool_controls_subjob = pool_applet.run(
{
"inputs": [ctl1_ta, ctl2_ta],
"prefix": "PL_ctls"
},
name='Pool controls')
pooled_controls = pool_controls_subjob.get_output_ref("pooled")
# always use the pooled controls for the pool
control_for_pool = pooled_controls
pool_ctl_msg = "pooled controls"
# use the pooled controls for the reps depending on the ratio of rep to
# control reads
ratio_ctl_reads = float(ntags_ctl1) / float(ntags_ctl2)
if ratio_ctl_reads < 1:
ratio_ctl_reads = 1 / ratio_ctl_reads
ratio_cutoff = 1.2
if ratio_ctl_reads > ratio_cutoff:
logger.info(
"Number of reads in controls differ by > factor of %f. Using pooled controls."
% (ratio_cutoff))
rep1_control = pooled_controls
rep2_control = pooled_controls
else:
if ntags_ctl1 < ntags_rep1:
logger.info(
"Fewer reads in control replicate 1 than experiment replicate 1. Using pooled controls for replicate 1."
)
rep1_control = pooled_controls
rep1_ctl_msg = "pooled controls"
elif not simplicate_experiment and ntags_ctl2 < ntags_rep2:
logger.info(
"Fewer reads in control replicate 2 than experiment replicate 2. Using pooled controls for replicate 2."
)
rep2_control = pooled_controls
rep2_ctl_msg = "pooled controls"
else:
logger.info("Using distinct controls for replicate 1 and 2.")
rep1_control = ctl1_ta # default. May be changed later.
rep2_control = ctl2_ta # default. May be changed later.
rep1_ctl_msg = "control rep1"
rep2_ctl_msg = "control rep2"
pseudoreplicator_applet = dxpy.find_one_data_object(
classname='applet',
name='pseudoreplicator',
zero_ok=False,
more_ok=False,
return_handler=True)
rep1_pr_subjob = pseudoreplicator_applet.run({"input_tags": rep1_ta})
if not simplicate_experiment:
rep2_pr_subjob = pseudoreplicator_applet.run({"input_tags": rep2_ta})
pool_pr1_subjob = pool_applet.run({
"inputs": [
rep1_pr_subjob.get_output_ref("pseudoreplicate1"),
rep2_pr_subjob.get_output_ref("pseudoreplicate1")
],
"prefix":
'PPR1'
})
pool_pr2_subjob = pool_applet.run({
"inputs": [
rep1_pr_subjob.get_output_ref("pseudoreplicate2"),
rep2_pr_subjob.get_output_ref("pseudoreplicate2")
],
"prefix":
'PPR2'
})
common_args = {
'chrom_sizes': chrom_sizes,
'genomesize': genomesize,
'narrowpeak_as': narrowpeak_as,
'gappedpeak_as': gappedpeak_as,
'broadpeak_as': broadpeak_as
}
# if the fragment_length argument is given, update macs2 input
if fragment_length is not None:
common_args.update({'fragment_length': fragment_length})
common_args.update({'prefix': 'r1'})
rep1_peaks_subjob = macs2(rep1_ta, rep1_control, rep1_xcor, **common_args)
common_args.update({'prefix': 'r1pr1'})
rep1pr1_peaks_subjob = macs2(
rep1_pr_subjob.get_output_ref("pseudoreplicate1"), rep1_control,
rep1_xcor, **common_args)
common_args.update({'prefix': 'r1pr2'})
rep1pr2_peaks_subjob = macs2(
rep1_pr_subjob.get_output_ref("pseudoreplicate2"), rep1_control,
rep1_xcor, **common_args)
if not simplicate_experiment:
common_args.update({'prefix': 'r2'})
rep2_peaks_subjob = macs2(rep2_ta, rep2_control, rep2_xcor,
**common_args)
common_args.update({'prefix': 'r2pr1'})
rep2pr1_peaks_subjob = macs2(
rep2_pr_subjob.get_output_ref("pseudoreplicate1"), rep2_control,
rep2_xcor, **common_args)
common_args.update({'prefix': 'r2pr2'})
rep2pr2_peaks_subjob = macs2(
rep2_pr_subjob.get_output_ref("pseudoreplicate2"), rep2_control,
rep2_xcor, **common_args)
common_args.update({'prefix': 'pool'})
pooled_peaks_subjob = macs2(
pooled_replicates, control_for_pool,
pooled_replicates_xcor_subjob.get_output_ref("CC_scores_file"),
**common_args)
common_args.update({'prefix': 'ppr1'})
pooledpr1_peaks_subjob = macs2(
pool_pr1_subjob.get_output_ref("pooled"), control_for_pool,
pooled_replicates_xcor_subjob.get_output_ref("CC_scores_file"),
**common_args)
common_args.update({'prefix': 'ppr2'})
pooledpr2_peaks_subjob = macs2(
pool_pr2_subjob.get_output_ref("pooled"), control_for_pool,
pooled_replicates_xcor_subjob.get_output_ref("CC_scores_file"),
**common_args)
output = {
'rep1_narrowpeaks':
rep1_peaks_subjob.get_output_ref("narrowpeaks"),
'rep1_gappedpeaks':
rep1_peaks_subjob.get_output_ref("gappedpeaks"),
'rep1_broadpeaks':
rep1_peaks_subjob.get_output_ref("broadpeaks"),
'rep1_narrowpeaks_bb':
rep1_peaks_subjob.get_output_ref("narrowpeaks_bb"),
'rep1_gappedpeaks_bb':
rep1_peaks_subjob.get_output_ref("gappedpeaks_bb"),
'rep1_broadpeaks_bb':
rep1_peaks_subjob.get_output_ref("broadpeaks_bb"),
'rep1_fc_signal':
rep1_peaks_subjob.get_output_ref("fc_signal"),
'rep1_pvalue_signal':
rep1_peaks_subjob.get_output_ref("pvalue_signal"),
'rep1pr1_narrowpeaks':
rep1pr1_peaks_subjob.get_output_ref("narrowpeaks"),
'rep1pr1_gappedpeaks':
rep1pr1_peaks_subjob.get_output_ref("gappedpeaks"),
'rep1pr1_broadpeaks':
rep1pr1_peaks_subjob.get_output_ref("broadpeaks"),
'rep1pr1_fc_signal':
rep1pr1_peaks_subjob.get_output_ref("fc_signal"),
'rep1pr1_pvalue_signal':
rep1pr1_peaks_subjob.get_output_ref("pvalue_signal"),
'rep1pr2_narrowpeaks':
rep1pr2_peaks_subjob.get_output_ref("narrowpeaks"),
'rep1pr2_gappedpeaks':
rep1pr2_peaks_subjob.get_output_ref("gappedpeaks"),
'rep1pr2_broadpeaks':
rep1pr2_peaks_subjob.get_output_ref("broadpeaks"),
'rep1pr2_fc_signal':
rep1pr2_peaks_subjob.get_output_ref("fc_signal"),
'rep1pr2_pvalue_signal':
rep1pr2_peaks_subjob.get_output_ref("pvalue_signal")
}
if not simplicate_experiment:
output.update({
'rep2_narrowpeaks':
rep2_peaks_subjob.get_output_ref("narrowpeaks"),
'rep2_gappedpeaks':
rep2_peaks_subjob.get_output_ref("gappedpeaks"),
'rep2_broadpeaks':
rep2_peaks_subjob.get_output_ref("broadpeaks"),
'rep2_narrowpeaks_bb':
rep2_peaks_subjob.get_output_ref("narrowpeaks_bb"),
'rep2_gappedpeaks_bb':
rep2_peaks_subjob.get_output_ref("gappedpeaks_bb"),
'rep2_broadpeaks_bb':
rep2_peaks_subjob.get_output_ref("broadpeaks_bb"),
'rep2_fc_signal':
rep2_peaks_subjob.get_output_ref("fc_signal"),
'rep2_pvalue_signal':
rep2_peaks_subjob.get_output_ref("pvalue_signal"),
'rep2pr1_narrowpeaks':
rep2pr1_peaks_subjob.get_output_ref("narrowpeaks"),
'rep2pr1_gappedpeaks':
rep2pr1_peaks_subjob.get_output_ref("gappedpeaks"),
'rep2pr1_broadpeaks':
rep2pr1_peaks_subjob.get_output_ref("broadpeaks"),
'rep2pr1_fc_signal':
rep2pr1_peaks_subjob.get_output_ref("fc_signal"),
'rep2pr1_pvalue_signal':
rep2pr1_peaks_subjob.get_output_ref("pvalue_signal"),
'rep2pr2_narrowpeaks':
rep2pr2_peaks_subjob.get_output_ref("narrowpeaks"),
'rep2pr2_gappedpeaks':
rep2pr2_peaks_subjob.get_output_ref("gappedpeaks"),
'rep2pr2_broadpeaks':
rep2pr2_peaks_subjob.get_output_ref("broadpeaks"),
'rep2pr2_fc_signal':
rep2pr2_peaks_subjob.get_output_ref("fc_signal"),
'rep2pr2_pvalue_signal':
rep2pr2_peaks_subjob.get_output_ref("pvalue_signal"),
'pooled_narrowpeaks':
pooled_peaks_subjob.get_output_ref("narrowpeaks"),
'pooled_gappedpeaks':
pooled_peaks_subjob.get_output_ref("gappedpeaks"),
'pooled_broadpeaks':
pooled_peaks_subjob.get_output_ref("broadpeaks"),
'pooled_narrowpeaks_bb':
pooled_peaks_subjob.get_output_ref("narrowpeaks_bb"),
'pooled_gappedpeaks_bb':
pooled_peaks_subjob.get_output_ref("gappedpeaks_bb"),
'pooled_broadpeaks_bb':
pooled_peaks_subjob.get_output_ref("broadpeaks_bb"),
'pooled_fc_signal':
pooled_peaks_subjob.get_output_ref("fc_signal"),
'pooled_pvalue_signal':
pooled_peaks_subjob.get_output_ref("pvalue_signal"),
'pooledpr1_narrowpeaks':
pooledpr1_peaks_subjob.get_output_ref("narrowpeaks"),
'pooledpr1_gappedpeaks':
pooledpr1_peaks_subjob.get_output_ref("gappedpeaks"),
'pooledpr1_broadpeaks':
pooledpr1_peaks_subjob.get_output_ref("broadpeaks"),
'pooledpr1_fc_signal':
pooledpr1_peaks_subjob.get_output_ref("fc_signal"),
'pooledpr1_pvalue_signal':
pooledpr1_peaks_subjob.get_output_ref("pvalue_signal"),
'pooledpr2_narrowpeaks':
pooledpr2_peaks_subjob.get_output_ref("narrowpeaks"),
'pooledpr2_gappedpeaks':
pooledpr2_peaks_subjob.get_output_ref("gappedpeaks"),
'pooledpr2_broadpeaks':
pooledpr2_peaks_subjob.get_output_ref("broadpeaks"),
'pooledpr2_fc_signal':
pooledpr2_peaks_subjob.get_output_ref("fc_signal"),
'pooledpr2_pvalue_signal':
pooledpr2_peaks_subjob.get_output_ref("pvalue_signal")
})
return output
def main(rep1_ta, ctl1_ta, rep1_xcor, rep1_paired_end,
npeaks, nodups, chrom_sizes, spp_version,
rep2_ta=None, ctl2_ta=None, rep2_xcor=None, rep2_paired_end=None,
as_file=None, idr_peaks=False, fragment_length=None, spp_instance=None):
rep1_ta_file = dxpy.DXFile(rep1_ta)
dxpy.download_dxfile(rep1_ta_file.get_id(), rep1_ta_file.name)
rep1_ta_filename = rep1_ta_file.name
ntags_rep1 = common.count_lines(rep1_ta_filename)
simplicate_experiment = rep1_ta and not rep2_ta
if simplicate_experiment:
logger.info("No rep2 tags specified so processing as a simplicate experiment.")
else:
logger.info("Rep1 and rep2 tags specified so processing as a replicated experiment.")
if not simplicate_experiment:
assert rep1_paired_end == rep2_paired_end, 'Mixed PE/SE not supported'
rep2_ta_file = dxpy.DXFile(rep2_ta)
dxpy.download_dxfile(rep2_ta_file.get_id(), rep2_ta_file.name)
rep2_ta_filename = rep2_ta_file.name
ntags_rep2 = common.count_lines(rep2_ta_filename)
paired_end = rep1_paired_end
unary_control = (ctl1_ta == ctl2_ta) or not ctl2_ta
ctl1_ta_file = dxpy.DXFile(ctl1_ta)
dxpy.download_dxfile(ctl1_ta_file.get_id(), ctl1_ta_file.name)
ctl1_ta_filename = ctl1_ta_file.name
if not unary_control:
ctl2_ta_file = dxpy.DXFile(ctl2_ta)
dxpy.download_dxfile(ctl2_ta_file.get_id(), ctl2_ta_file.name)
ctl2_ta_filename = ctl2_ta_file.name
else:
ctl2_ta_file = ctl1_ta_file
ctl2_ta_filename = ctl1_ta_file.name
ntags_ctl1 = common.count_lines(ctl1_ta_filename)
ntags_ctl2 = common.count_lines(ctl2_ta_filename)
rep1_control = ctl1_ta # default. May be changed later.
rep1_ctl_msg = "control rep1"
rep2_control = ctl2_ta # default. May be changed later.
rep2_ctl_msg = "control rep2"
rep_info = [(ntags_rep1, 'replicate 1', rep1_ta_filename)]
if not simplicate_experiment:
rep_info.append((ntags_rep2, 'replicate 2', rep2_ta_filename))
rep_info.extend(
[(ntags_ctl1, 'control 1', ctl1_ta_filename),
(ntags_ctl2, 'control 2', ctl2_ta_filename)])
for n, name, filename in rep_info:
logger.info("Found %d tags in %s file %s" % (n, name, filename))
subprocess.check_output('ls -l', shell=True, stderr=subprocess.STDOUT)
if not simplicate_experiment:
pool_applet = dxpy.find_one_data_object(
classname='applet',
name='pool',
project=dxpy.PROJECT_CONTEXT_ID,
zero_ok=False,
more_ok=False,
return_handler=True)
pool_replicates_subjob = \
pool_applet.run(
{"inputs": [rep1_ta, rep2_ta],
"prefix": 'pooled_reps'},
name='Pool replicates')
pooled_replicates = pool_replicates_subjob.get_output_ref("pooled")
pooled_replicates_xcor_subjob = \
xcor_only(
pooled_replicates,
paired_end,
spp_version,
name='Pool cross-correlation')
if unary_control:
logger.info("Only one control supplied.")
if not simplicate_experiment:
logger.info("Using one control for both replicate 1 and 2 and for the pool.")
rep2_control = rep1_control
control_for_pool = rep1_control
pool_ctl_msg = "one control"
else:
pool_controls_subjob = pool_applet.run(
{"inputs": [ctl1_ta, ctl2_ta],
"prefix": "PL_ctls"},
name='Pool controls')
pooled_controls = pool_controls_subjob.get_output_ref("pooled")
# always use the pooled controls for the pool
control_for_pool = pooled_controls
pool_ctl_msg = "pooled controls"
# use the pooled controls for the reps depending on the ratio of rep to
# control reads
ratio_ctl_reads = float(ntags_ctl1)/float(ntags_ctl2)
if ratio_ctl_reads < 1:
ratio_ctl_reads = 1/ratio_ctl_reads
ratio_cutoff = 1.2
if ratio_ctl_reads > ratio_cutoff:
logger.info(
"Number of reads in controls differ by > factor of %f. Using pooled controls."
% (ratio_cutoff))
rep1_control = pooled_controls
rep2_control = pooled_controls
else:
if ntags_ctl1 < ntags_rep1:
logger.info("Fewer reads in control replicate 1 than experiment replicate 1. Using pooled controls for replicate 1.")
rep1_control = pooled_controls
rep1_ctl_msg = "pooled controls"
elif not simplicate_experiment and ntags_ctl2 < ntags_rep2:
logger.info("Fewer reads in control replicate 2 than experiment replicate 2. Using pooled controls for replicate 2.")
rep2_control = pooled_controls
rep2_ctl_msg = "pooled controls"
else:
logger.info(
"Using distinct controls for replicate 1 and 2.")
rep1_control = ctl1_ta # default. May be changed later.
rep2_control = ctl2_ta # default. May be changed later.
rep1_ctl_msg = "control rep1"
rep2_ctl_msg = "control rep2"
common_args = {
'chrom_sizes': chrom_sizes,
'spp_version': spp_version,
'as_file': as_file,
'spp_instance': spp_instance
}
if fragment_length is not None:
common_args.update({'fragment_length': fragment_length})
rep1_peaks_subjob = spp(
rep1_ta,
rep1_control,
rep1_xcor,
bigbed=True,
name='Rep1 peaks vs %s' % (rep1_ctl_msg),
prefix='R1', **common_args)
if not simplicate_experiment:
rep2_peaks_subjob = spp(
rep2_ta,
rep2_control,
rep2_xcor,
bigbed=True,
name='Rep2 peaks vs %s' % (rep2_ctl_msg),
prefix='R2', **common_args)
pooled_peaks_subjob = spp(
pooled_replicates,
control_for_pool,
pooled_replicates_xcor_subjob.get_output_ref("CC_scores_file"),
bigbed=True,
name='Pooled peaks vs %s' % (pool_ctl_msg),
prefix='PL', **common_args)
output = {
'rep1_peaks': rep1_peaks_subjob.get_output_ref("peaks"),
'rep1_peaks_bb': rep1_peaks_subjob.get_output_ref("peaks_bb"),
'rep1_xcor_plot': rep1_peaks_subjob.get_output_ref("xcor_plot"),
'rep1_xcor_scores': rep1_peaks_subjob.get_output_ref("xcor_scores")
}
if not simplicate_experiment:
output.update({
'rep2_peaks': rep2_peaks_subjob.get_output_ref("peaks"),
'rep2_peaks_bb': rep2_peaks_subjob.get_output_ref("peaks_bb"),
'rep2_xcor_plot': rep2_peaks_subjob.get_output_ref("xcor_plot"),
'rep2_xcor_scores': rep2_peaks_subjob.get_output_ref("xcor_scores"),
'pooled_peaks': pooled_peaks_subjob.get_output_ref("peaks"),
'pooled_peaks_bb': pooled_peaks_subjob.get_output_ref("peaks_bb"),
'pooled_xcor_plot': pooled_peaks_subjob.get_output_ref("xcor_plot"),
'pooled_xcor_scores': pooled_peaks_subjob.get_output_ref("xcor_scores")
})
if idr_peaks: # also call peaks on pseudoreplicates for IDR
pseudoreplicator_applet = \
dxpy.find_one_data_object(
classname='applet',
name='pseudoreplicator',
project=dxpy.PROJECT_CONTEXT_ID,
zero_ok=False,
more_ok=False,
return_handler=True)
rep1_pr_subjob = \
pseudoreplicator_applet.run(
{"input_tags": rep1_ta,
"prefix": 'R1PR'},
name='Pseudoreplicate rep1 -> R1PR1,2')
rep1pr1_peaks_subjob = spp(
rep1_pr_subjob.get_output_ref("pseudoreplicate1"),
rep1_control,
rep1_xcor,
bigbed=False,
name='R1PR1 peaks vs %s' % (rep1_ctl_msg),
prefix='R1PR1', **common_args)
rep1pr2_peaks_subjob = spp(
rep1_pr_subjob.get_output_ref("pseudoreplicate2"),
rep1_control,
rep1_xcor,
bigbed=False,
name='R1PR2 peaks vs %s' % (rep1_ctl_msg),
prefix='R1PR2', **common_args)
output.update({
'rep1pr1_peaks': rep1pr1_peaks_subjob.get_output_ref("peaks"),
'rep1pr2_peaks': rep1pr2_peaks_subjob.get_output_ref("peaks")
})
if not simplicate_experiment:
rep2_pr_subjob = \
pseudoreplicator_applet.run(
{"input_tags": rep2_ta,
"prefix": 'R2PR'},
name='Pseudoreplicate rep2 -> R2PR1,2')
pool_pr1_subjob = pool_applet.run({
"inputs": [
rep1_pr_subjob.get_output_ref("pseudoreplicate1"),
rep2_pr_subjob.get_output_ref("pseudoreplicate1")],
"prefix": 'PPR1'},
name='Pool R1PR1+R2PR1 -> PPR1')
pool_pr2_subjob = pool_applet.run({
"inputs": [
rep1_pr_subjob.get_output_ref("pseudoreplicate2"),
rep2_pr_subjob.get_output_ref("pseudoreplicate2")],
"prefix": 'PPR2'},
name='Pool R1PR2+R2PR2 -> PPR2')
rep2pr1_peaks_subjob = spp(
rep2_pr_subjob.get_output_ref("pseudoreplicate1"),
rep2_control,
rep2_xcor,
bigbed=False,
name='R2PR1 peaks vs %s' % (rep2_ctl_msg),
prefix='R2PR1', **common_args)
rep2pr2_peaks_subjob = spp(
rep2_pr_subjob.get_output_ref("pseudoreplicate2"),
rep2_control,
rep2_xcor,
bigbed=False,
name='R2PR2 peaks vs %s' % (rep2_ctl_msg),
prefix='R2PR2', **common_args)
pooledpr1_peaks_subjob = spp(
pool_pr1_subjob.get_output_ref("pooled"),
control_for_pool,
pooled_replicates_xcor_subjob.get_output_ref("CC_scores_file"),
bigbed=False,
name='PPR1 peaks vs %s' % (pool_ctl_msg),
prefix='PPR1', **common_args)
pooledpr2_peaks_subjob = spp(
pool_pr2_subjob.get_output_ref("pooled"),
control_for_pool,
pooled_replicates_xcor_subjob.get_output_ref("CC_scores_file"),
bigbed=False,
name='PPR2 peaks vs %s' % (pool_ctl_msg),
prefix='PPR2', **common_args)
output.update({
'rep2pr1_peaks': rep2pr1_peaks_subjob.get_output_ref("peaks"),
'rep2pr2_peaks': rep2pr2_peaks_subjob.get_output_ref("peaks"),
'pooledpr1_peaks': pooledpr1_peaks_subjob.get_output_ref("peaks"),
'pooledpr2_peaks': pooledpr2_peaks_subjob.get_output_ref("peaks"),
})
return output
def main(experiment, control, xcor_scores_input, npeaks, nodups, bigbed,
chrom_sizes, spp_version, as_file=None, prefix=None):
# The following line(s) initialize your data object inputs on the platform
# into dxpy.DXDataObject instances that you can start using immediately.
experiment_file = dxpy.DXFile(experiment)
control_file = dxpy.DXFile(control)
xcor_scores_input_file = dxpy.DXFile(xcor_scores_input)
chrom_sizes_file = dxpy.DXFile(chrom_sizes)
chrom_sizes_filename = chrom_sizes_file.name
dxpy.download_dxfile(chrom_sizes_file.get_id(), chrom_sizes_filename)
if bigbed:
as_file_file = dxpy.DXFile(as_file)
as_file_filename = as_file_file.name
dxpy.download_dxfile(as_file_file.get_id(), as_file_filename)
# The following line(s) download your file inputs to the local file system
# using variable names for the filenames.
experiment_filename = experiment_file.name
dxpy.download_dxfile(experiment_file.get_id(), experiment_filename)
control_filename = control_file.name
dxpy.download_dxfile(control_file.get_id(), control_filename)
xcor_scores_input_filename = xcor_scores_input_file.name
dxpy.download_dxfile(
xcor_scores_input_file.get_id(), xcor_scores_input_filename)
if not prefix:
output_filename_prefix = \
experiment_filename.rstrip('.gz').rstrip('.tagAlign')
else:
output_filename_prefix = prefix
peaks_filename = output_filename_prefix + '.regionPeak'
# spp adds .gz, so this is the file name that's actually created
final_peaks_filename = peaks_filename + '.gz'
xcor_plot_filename = output_filename_prefix + '.pdf'
xcor_scores_filename = output_filename_prefix + '.ccscores'
logger.info(subprocess.check_output(
'ls -l', shell=True, stderr=subprocess.STDOUT))
# third column in the cross-correlation scores input file
fraglen_column = 3
with open(xcor_scores_input_filename, 'r') as f:
line = f.readline()
fragment_length = int(line.split('\t')[fraglen_column-1])
logger.info("Read fragment length: %d" % (fragment_length))
spp_tarball = SPP_VERSION_MAP.get(spp_version)
assert spp_tarball, "spp version %s is not supported" % (spp_version)
if nodups:
run_spp = '/phantompeakqualtools/run_spp_nodups.R'
else:
run_spp = '/phantompeakqualtools/run_spp.R'
# install spp
subprocess.check_output(shlex.split('R CMD INSTALL %s' % (spp_tarball)))
spp_command = (
"Rscript %s -p=%d -c=%s -i=%s -npeak=%d -speak=%d -savr=%s -savp=%s -rf -out=%s"
% (run_spp, cpu_count(), experiment_filename, control_filename, npeaks,
fragment_length, peaks_filename, xcor_plot_filename,
xcor_scores_filename))
logger.info(spp_command)
subprocess.check_call(shlex.split(spp_command))
# when one of the peak coordinates are an exact multiple of 10, spp (R)
# outputs the coordinate in scientific notation
# this changes any such coodinates to decimal notation
# this assumes 10-column output and that the 2nd and 3rd columns are
# coordinates
# the ($2>0)?$2:0) is needed because spp sometimes calls peaks with a
# negative start coordinate (particularly chrM) and will cause slopBed
# to halt at that line, truncating the output of the pipe
# slopBed adjusts feature end coordinates that go off the end of the
# chromosome
# bedClip removes any features that are still not within the boundaries of
# the chromosome
fix_coordinate_peaks_filename = \
output_filename_prefix + '.fixcoord.regionPeak'
out, err = common.run_pipe([
"gzip -dc %s" % (final_peaks_filename),
"tee %s" % (peaks_filename),
r"""awk 'BEGIN{OFS="\t"}{print $1,sprintf("%i",($2>0)?$2:0),sprintf("%i",$3),$4,$5,$6,$7,$8,$9,$10}'""",
'slopBed -i stdin -g %s -b 0' % (chrom_sizes_filename),
'bedClip stdin %s %s' % (chrom_sizes_filename, fix_coordinate_peaks_filename)
])
# These lines transfer the peaks files to the temporary workspace for
# debugging later
# Only at the end are the final files uploaded that will be returned from
# the applet
dxpy.upload_local_file(peaks_filename)
dxpy.upload_local_file(fix_coordinate_peaks_filename)
n_spp_peaks = common.count_lines(peaks_filename)
logger.info("%s peaks called by spp" % (n_spp_peaks))
logger.info(
"%s of those peaks removed due to bad coordinates"
% (n_spp_peaks - common.count_lines(fix_coordinate_peaks_filename)))
print("First 50 peaks")
subprocess.check_output(
'head -50 %s' % (fix_coordinate_peaks_filename),
shell=True)
if bigbed:
peaks_bb_filename = \
common.bed2bb(fix_coordinate_peaks_filename, chrom_sizes_filename, as_file_filename)
if peaks_bb_filename:
peaks_bb = dxpy.upload_local_file(peaks_bb_filename)
if not filecmp.cmp(peaks_filename,fix_coordinate_peaks_filename):
logger.info("Returning peaks with fixed coordinates")
subprocess.check_call(shlex.split('gzip -n %s' % (fix_coordinate_peaks_filename)))
final_peaks_filename = fix_coordinate_peaks_filename + '.gz'
subprocess.check_call('ls -l', shell=True)
# print subprocess.check_output('head %s' %(final_peaks_filename), shell=True, stderr=subprocess.STDOUT)
# print subprocess.check_output('head %s' %(xcor_scores_filename), shell=True, stderr=subprocess.STDOUT)
peaks = dxpy.upload_local_file(final_peaks_filename)
xcor_plot = dxpy.upload_local_file(xcor_plot_filename)
xcor_scores = dxpy.upload_local_file(xcor_scores_filename)
output = {}
output["peaks"] = dxpy.dxlink(peaks)
output["xcor_plot"] = dxpy.dxlink(xcor_plot)
output["xcor_scores"] = dxpy.dxlink(xcor_scores)
if bigbed and peaks_bb_filename:
output["peaks_bb"] = dxpy.dxlink(peaks_bb)
return output
def main(rep1_ta, rep2_ta, ctl1_ta, ctl2_ta, rep1_xcor, rep2_xcor,
rep1_paired_end, rep2_paired_end, chrom_sizes, genomesize,
narrowpeak_as, gappedpeak_as, broadpeak_as):
assert rep1_paired_end == rep2_paired_end, 'Mixed PE/SE not supported'
paired_end = rep1_paired_end
rep1_ta_file = dxpy.DXFile(rep1_ta)
rep2_ta_file = dxpy.DXFile(rep2_ta)
unary_control = ctl1_ta == ctl2_ta
ctl1_ta_file = dxpy.DXFile(ctl1_ta)
ctl2_ta_file = dxpy.DXFile(ctl2_ta)
# not necessary to actually download these - just pass through
# rep1_xcor_file = dxpy.DXFile(rep1_xcor)
# rep2_xcor_file = dxpy.DXFile(rep2_xcor)
dxpy.download_dxfile(rep1_ta_file.get_id(), rep1_ta_file.name)
dxpy.download_dxfile(rep2_ta_file.get_id(), rep2_ta_file.name)
dxpy.download_dxfile(ctl1_ta_file.get_id(), ctl1_ta_file.name)
dxpy.download_dxfile(ctl2_ta_file.get_id(), ctl2_ta_file.name)
# not necessary to actually download these - just pass through
# dxpy.download_dxfile(rep1_xcor_file.get_id(), rep1_xcor_file.name)
# dxpy.download_dxfile(rep2_xcor_file.get_id(), rep2_xcor_file.name)
rep1_ta_filename = rep1_ta_file.name
rep2_ta_filename = rep2_ta_file.name
ctl1_ta_filename = ctl1_ta_file.name
ctl2_ta_filename = ctl2_ta_file.name
# not necessary to actually download these - just pass through
# rep1_xcor_filename = rep1_xcor_file.name
# rep2_xcor_filename = rep2_xcor_file.name
ntags_rep1 = common.count_lines(rep1_ta_filename)
ntags_rep2 = common.count_lines(rep2_ta_filename)
ntags_ctl1 = common.count_lines(ctl1_ta_filename)
ntags_ctl2 = common.count_lines(ctl2_ta_filename)
for n, name, filename in [(ntags_rep1, 'replicate 1', rep1_ta_filename),
(ntags_rep2, 'replicate 2', rep2_ta_filename),
(ntags_ctl1, 'control 1', ctl1_ta_filename),
(ntags_ctl2, 'control 2', ctl2_ta_filename)]:
logger.info("Found %d tags in %s file %s" % (n, name, filename))
subprocess.check_output('ls -l', shell=True, stderr=subprocess.STDOUT)
pool_applet = dxpy.find_one_data_object(
classname='applet',
name='pool',
zero_ok=False,
more_ok=False,
return_handler=True)
pool_replicates_subjob = pool_applet.run({
"inputs": [rep1_ta, rep2_ta],
"prefix": "PL_reps"})
pooled_replicates = pool_replicates_subjob.get_output_ref("pooled")
rep1_control = ctl1_ta # default. May be changed later.
rep2_control = ctl2_ta # default. May be changed later.
if unary_control:
logger.info("Only one control supplied. Using it for both replicate 1 and 2 and for the pool.")
control_for_pool = rep1_control
else:
pool_controls_subjob = pool_applet.run({
"inputs": [ctl1_ta, ctl2_ta],
"prefix": "PL_ctls"})
pooled_controls = pool_controls_subjob.get_output_ref("pooled")
# always use the pooled controls for the pool
control_for_pool = pooled_controls
# use the pooled controls for the reps depending on the ratio of rep to
# control reads
ratio_ctl_reads = float(ntags_ctl1)/float(ntags_ctl2)
if ratio_ctl_reads < 1:
ratio_ctl_reads = 1/ratio_ctl_reads
ratio_cutoff = 1.2
if ratio_ctl_reads > ratio_cutoff:
logger.info(
"Number of reads in controls differ by > factor of %f. Using pooled controls."
% (ratio_cutoff))
rep1_control = pooled_controls
rep2_control = pooled_controls
else:
if ntags_ctl1 < ntags_rep1:
logger.info("Fewer reads in control replicate 1 than experiment replicate 1. Using pooled controls for replicate 1.")
rep1_control = pooled_controls
elif ntags_ctl2 < ntags_rep2:
logger.info("Fewer reads in control replicate 2 than experiment replicate 2. Using pooled controls for replicate 2.")
rep2_control = pooled_controls
else:
logger.info(
"Using distinct controls for replicate 1 and 2.")
pseudoreplicator_applet = dxpy.find_one_data_object(
classname='applet',
name='pseudoreplicator',
zero_ok=False,
more_ok=False,
return_handler=True)
rep1_pr_subjob = pseudoreplicator_applet.run({"input_tags": rep1_ta})
rep2_pr_subjob = pseudoreplicator_applet.run({"input_tags": rep2_ta})
pool_pr1_subjob = pool_applet.run(
{"inputs": [rep1_pr_subjob.get_output_ref("pseudoreplicate1"),
rep2_pr_subjob.get_output_ref("pseudoreplicate1")],
"prefix": 'PPR1'})
pool_pr2_subjob = pool_applet.run(
{"inputs": [rep1_pr_subjob.get_output_ref("pseudoreplicate2"),
rep2_pr_subjob.get_output_ref("pseudoreplicate2")],
"prefix": 'PPR2'})
pooled_replicates_xcor_subjob = xcor_only(pooled_replicates, paired_end)
# no longer calculated - now we take the cross-correlation metrics for the
# pseudoreplicates as those from the true reps
# rep1_pr1_xcor_subjob = \
# xcor_only(rep1_pr_subjob.get_output_ref("pseudoreplicate1"), paired_end)
# rep1_pr2_xcor_subjob = \
# xcor_only(rep1_pr_subjob.get_output_ref("pseudoreplicate2"), paired_end)
# rep2_pr1_xcor_subjob = \
# xcor_only(rep2_pr_subjob.get_output_ref("pseudoreplicate1"), paired_end)
# rep2_pr2_xcor_subjob = \
# xcor_only(rep2_pr_subjob.get_output_ref("pseudoreplicate2"), paired_end)
# pool_pr1_xcor_subjob = \
# xcor_only(pool_pr1_subjob.get_output_ref("pooled"), paired_end)
# pool_pr2_xcor_subjob = \
# xcor_only(pool_pr2_subjob.get_output_ref("pooled"), paired_end)
common_args = {
'chrom_sizes': chrom_sizes,
'genomesize': genomesize,
'narrowpeak_as': narrowpeak_as,
'gappedpeak_as': gappedpeak_as,
'broadpeak_as': broadpeak_as
}
common_args.update({'prefix': 'r1'})
rep1_peaks_subjob = macs2( rep1_ta,
rep1_control,
rep1_xcor, **common_args)
common_args.update({'prefix': 'r2'})
rep2_peaks_subjob = macs2( rep2_ta,
rep2_control,
rep2_xcor, **common_args)
common_args.update({'prefix': 'pool'})
pooled_peaks_subjob = macs2( pooled_replicates,
control_for_pool,
pooled_replicates_xcor_subjob.get_output_ref("CC_scores_file"), **common_args)
common_args.update({'prefix': 'r1pr1'})
rep1pr1_peaks_subjob = macs2( rep1_pr_subjob.get_output_ref("pseudoreplicate1"),
rep1_control,
rep1_xcor, **common_args)
common_args.update({'prefix': 'r1pr2'})
rep1pr2_peaks_subjob = macs2( rep1_pr_subjob.get_output_ref("pseudoreplicate2"),
rep1_control,
rep1_xcor, **common_args)
common_args.update({'prefix': 'r2pr1'})
rep2pr1_peaks_subjob = macs2( rep2_pr_subjob.get_output_ref("pseudoreplicate1"),
rep2_control,
rep2_xcor, **common_args)
common_args.update({'prefix': 'r2pr2'})
rep2pr2_peaks_subjob = macs2( rep2_pr_subjob.get_output_ref("pseudoreplicate2"),
rep2_control,
rep2_xcor, **common_args)
common_args.update({'prefix': 'ppr1'})
pooledpr1_peaks_subjob = macs2( pool_pr1_subjob.get_output_ref("pooled"),
control_for_pool,
pooled_replicates_xcor_subjob.get_output_ref("CC_scores_file"), **common_args)
common_args.update({'prefix': 'ppr2'})
pooledpr2_peaks_subjob = macs2( pool_pr2_subjob.get_output_ref("pooled"),
control_for_pool,
pooled_replicates_xcor_subjob.get_output_ref("CC_scores_file"), **common_args)
output = {
'rep1_narrowpeaks': rep1_peaks_subjob.get_output_ref("narrowpeaks"),
'rep1_gappedpeaks': rep1_peaks_subjob.get_output_ref("gappedpeaks"),
'rep1_broadpeaks': rep1_peaks_subjob.get_output_ref("broadpeaks"),
'rep1_narrowpeaks_bb': rep1_peaks_subjob.get_output_ref("narrowpeaks_bb"),
'rep1_gappedpeaks_bb': rep1_peaks_subjob.get_output_ref("gappedpeaks_bb"),
'rep1_broadpeaks_bb': rep1_peaks_subjob.get_output_ref("broadpeaks_bb"),
'rep1_fc_signal': rep1_peaks_subjob.get_output_ref("fc_signal"),
'rep1_pvalue_signal': rep1_peaks_subjob.get_output_ref("pvalue_signal"),
'rep2_narrowpeaks': rep2_peaks_subjob.get_output_ref("narrowpeaks"),
'rep2_gappedpeaks': rep2_peaks_subjob.get_output_ref("gappedpeaks"),
'rep2_broadpeaks': rep2_peaks_subjob.get_output_ref("broadpeaks"),
'rep2_narrowpeaks_bb': rep2_peaks_subjob.get_output_ref("narrowpeaks_bb"),
'rep2_gappedpeaks_bb': rep2_peaks_subjob.get_output_ref("gappedpeaks_bb"),
'rep2_broadpeaks_bb': rep2_peaks_subjob.get_output_ref("broadpeaks_bb"),
'rep2_fc_signal': rep2_peaks_subjob.get_output_ref("fc_signal"),
'rep2_pvalue_signal': rep2_peaks_subjob.get_output_ref("pvalue_signal"),
'pooled_narrowpeaks': pooled_peaks_subjob.get_output_ref("narrowpeaks"),
'pooled_gappedpeaks': pooled_peaks_subjob.get_output_ref("gappedpeaks"),
'pooled_broadpeaks': pooled_peaks_subjob.get_output_ref("broadpeaks"),
'pooled_narrowpeaks_bb': pooled_peaks_subjob.get_output_ref("narrowpeaks_bb"),
'pooled_gappedpeaks_bb': pooled_peaks_subjob.get_output_ref("gappedpeaks_bb"),
'pooled_broadpeaks_bb': pooled_peaks_subjob.get_output_ref("broadpeaks_bb"),
'pooled_fc_signal': pooled_peaks_subjob.get_output_ref("fc_signal"),
'pooled_pvalue_signal': pooled_peaks_subjob.get_output_ref("pvalue_signal"),
'rep1pr1_narrowpeaks': rep1pr1_peaks_subjob.get_output_ref("narrowpeaks"),
'rep1pr1_gappedpeaks': rep1pr1_peaks_subjob.get_output_ref("gappedpeaks"),
'rep1pr1_broadpeaks': rep1pr1_peaks_subjob.get_output_ref("broadpeaks"),
'rep1pr1_fc_signal': rep1pr1_peaks_subjob.get_output_ref("fc_signal"),
'rep1pr1_pvalue_signal': rep1pr1_peaks_subjob.get_output_ref("pvalue_signal"),
'rep1pr2_narrowpeaks': rep1pr2_peaks_subjob.get_output_ref("narrowpeaks"),
'rep1pr2_gappedpeaks': rep1pr2_peaks_subjob.get_output_ref("gappedpeaks"),
'rep1pr2_broadpeaks': rep1pr2_peaks_subjob.get_output_ref("broadpeaks"),
'rep1pr2_fc_signal': rep1pr2_peaks_subjob.get_output_ref("fc_signal"),
'rep1pr2_pvalue_signal': rep1pr2_peaks_subjob.get_output_ref("pvalue_signal"),
'rep2pr1_narrowpeaks': rep2pr1_peaks_subjob.get_output_ref("narrowpeaks"),
'rep2pr1_gappedpeaks': rep2pr1_peaks_subjob.get_output_ref("gappedpeaks"),
'rep2pr1_broadpeaks': rep2pr1_peaks_subjob.get_output_ref("broadpeaks"),
'rep2pr1_fc_signal': rep2pr1_peaks_subjob.get_output_ref("fc_signal"),
'rep2pr1_pvalue_signal': rep2pr1_peaks_subjob.get_output_ref("pvalue_signal"),
'rep2pr2_narrowpeaks': rep2pr2_peaks_subjob.get_output_ref("narrowpeaks"),
'rep2pr2_gappedpeaks': rep2pr2_peaks_subjob.get_output_ref("gappedpeaks"),
'rep2pr2_broadpeaks': rep2pr2_peaks_subjob.get_output_ref("broadpeaks"),
'rep2pr2_fc_signal': rep2pr2_peaks_subjob.get_output_ref("fc_signal"),
'rep2pr2_pvalue_signal': rep2pr2_peaks_subjob.get_output_ref("pvalue_signal"),
'pooledpr1_narrowpeaks': pooledpr1_peaks_subjob.get_output_ref("narrowpeaks"),
'pooledpr1_gappedpeaks': pooledpr1_peaks_subjob.get_output_ref("gappedpeaks"),
'pooledpr1_broadpeaks': pooledpr1_peaks_subjob.get_output_ref("broadpeaks"),
'pooledpr1_fc_signal': pooledpr1_peaks_subjob.get_output_ref("fc_signal"),
'pooledpr1_pvalue_signal': pooledpr1_peaks_subjob.get_output_ref("pvalue_signal"),
'pooledpr2_narrowpeaks': pooledpr2_peaks_subjob.get_output_ref("narrowpeaks"),
'pooledpr2_gappedpeaks': pooledpr2_peaks_subjob.get_output_ref("gappedpeaks"),
'pooledpr2_broadpeaks': pooledpr2_peaks_subjob.get_output_ref("broadpeaks"),
'pooledpr2_fc_signal': pooledpr2_peaks_subjob.get_output_ref("fc_signal"),
'pooledpr2_pvalue_signal': pooledpr2_peaks_subjob.get_output_ref("pvalue_signal")
}
return output
def main(experiment, reps_peaks, r1pr_peaks, r2pr_peaks, pooledpr_peaks, chrom_sizes, as_file, blacklist=None):
#TODO for now just taking the peak files. This applet should actually call IDR instead of
#putting that in the workflow populator script
# Initialize the data object inputs on the platform into
# dxpy.DXDataObject instances.
reps_peaks_file = dxpy.DXFile(reps_peaks)
r1pr_peaks_file = dxpy.DXFile(r1pr_peaks)
r2pr_peaks_file = dxpy.DXFile(r2pr_peaks)
pooledpr_peaks_file = dxpy.DXFile(pooledpr_peaks)
chrom_sizes_file = dxpy.DXFile(chrom_sizes)
as_file_file = dxpy.DXFile(as_file)
if blacklist is not None:
blacklist_file = dxpy.DXFile(blacklist)
blacklist_filename = 'blacklist_%s' %(blacklist_file.name)
dxpy.download_dxfile(blacklist_file.get_id(), blacklist_filename)
blacklist_filename = common.uncompress(blacklist_filename)
# Download the file inputs to the local file system.
#Need to prepend something to ensure the local filenames will be unique
reps_peaks_filename = 'true_%s' %(reps_peaks_file.name)
r1pr_peaks_filename = 'r1pr_%s' %(r1pr_peaks_file.name)
r2pr_peaks_filename = 'r2pr_%s' %(r2pr_peaks_file.name)
pooledpr_peaks_filename = 'pooledpr_%s' %(pooledpr_peaks_file.name)
chrom_sizes_filename = chrom_sizes_file.name
as_file_filename = as_file_file.name
dxpy.download_dxfile(reps_peaks_file.get_id(), reps_peaks_filename)
dxpy.download_dxfile(r1pr_peaks_file.get_id(), r1pr_peaks_filename)
dxpy.download_dxfile(r2pr_peaks_file.get_id(), r2pr_peaks_filename)
dxpy.download_dxfile(pooledpr_peaks_file.get_id(), pooledpr_peaks_filename)
dxpy.download_dxfile(chrom_sizes_file.get_id(), chrom_sizes_filename)
dxpy.download_dxfile(as_file_file.get_id(), as_file_filename)
print subprocess.check_output('ls -l', shell=True)
reps_peaks_filename = common.uncompress(reps_peaks_filename)
r1pr_peaks_filename = common.uncompress(r1pr_peaks_filename)
r2pr_peaks_filename = common.uncompress(r2pr_peaks_filename)
pooledpr_peaks_filename = common.uncompress(pooledpr_peaks_filename)
Nt = common.count_lines(reps_peaks_filename)
print "%d peaks from true replicates" %(Nt)
N1 = common.count_lines(r1pr_peaks_filename)
print "%d peaks from rep1 self-pseudoreplicates" %(N1)
N2 = common.count_lines(r2pr_peaks_filename)
print "%d peaks from rep2 self-pseudoreplicates" %(N2)
Np = common.count_lines(pooledpr_peaks_filename)
print "%d peaks from pooled pseudoreplicates" %(Np)
conservative_set_filename = '%s_final_conservative.narrowPeak' %(experiment)
if blacklist is not None:
blacklist_filter(reps_peaks_filename, conservative_set_filename, blacklist_filename)
else:
conservative_set_filename = reps_peaks_filename
Ncb = common.count_lines(conservative_set_filename)
print "%d peaks blacklisted from the conservative set" %(Nt-Ncb)
if Nt >= Np:
peaks_to_filter_filename = reps_peaks_filename
No = Nt
else:
peaks_to_filter_filename = pooledpr_peaks_filename
No = Np
optimal_set_filename = '%s_final_optimal.narrowPeak' %(experiment)
if blacklist is not None:
blacklist_filter(peaks_to_filter_filename, optimal_set_filename, blacklist_filename)
else:
optimal_set_filename = peaks_to_filter_filename
Nob = common.count_lines(optimal_set_filename)
print "%d peaks blacklisted from the optimal set" %(No-Nob)
rescue_ratio = float(max(Np,Nt)) / float(min(Np,Nt))
self_consistency_ratio = float(max(N1,N2)) / float(min(N1,N2))
if rescue_ratio > 2 and self_consistency_ratio > 2:
reproducibility = 'fail'
elif rescue_ratio > 2 or self_consistency_ratio > 2:
reproducibility = 'borderline'
else:
reproducibility = 'pass'
output = {}
#bedtobigbed often fails, so skip creating the bb if it does
conservative_set_bb_filename = common.bed2bb(conservative_set_filename, chrom_sizes_filename, as_file_filename)
optimal_set_bb_filename = common.bed2bb(optimal_set_filename, chrom_sizes_filename, as_file_filename)
if conservative_set_bb_filename:
conservative_set_bb_output = dxpy.upload_local_file(conservative_set_bb_filename)
output.update({"conservative_set_bb": dxpy.dxlink(conservative_set_bb_output)})
if optimal_set_bb_filename:
optimal_set_bb_output = dxpy.upload_local_file(optimal_set_bb_filename)
output.update({"optimal_set_bb": dxpy.dxlink(optimal_set_bb_output)})
output.update({
"Nt": Nt,
"N1": N1,
"N2": N2,
"Np": Np,
"conservative_set": dxpy.dxlink(dxpy.upload_local_file(common.compress(conservative_set_filename))),
"optimal_set": dxpy.dxlink(dxpy.upload_local_file(common.compress(optimal_set_filename))),
"rescue_ratio": rescue_ratio,
"self_consistency_ratio": self_consistency_ratio,
"reproducibility_test": reproducibility,
"No": Nob,
"Nc": Ncb
})
logging.info("Exiting with output: %s", output)
return output
def process(self):
'''
#find pooled peaks that are in (rep1 AND rep2)
out, err = common.run_pipe([
'intersectBed -wa -f 0.50 -r -a %s -b %s' %(pooled_peaks_fn, rep1_peaks_fn),
'intersectBed -wa -f 0.50 -r -a stdin -b %s' %(rep2_peaks_fn)
], overlap_tr_fn)
print "%d peaks overlap with both true replicates" %(common.count_lines(overlap_tr_fn))
#pooled peaks that are in (pooledpseudorep1 AND pooledpseudorep2)
out, err = common.run_pipe([
'intersectBed -wa -f 0.50 -r -a %s -b %s' %(pooled_peaks_fn, pooledpr1_peaks_fn),
'intersectBed -wa -f 0.50 -r -a stdin -b %s' %(pooledpr2_peaks_fn)
], overlap_pr_fn)
print "%d peaks overlap with both pooled pseudoreplicates" %(common.count_lines(overlap_pr_fn))
#combined pooled peaks in (rep1 AND rep2) OR (pooledpseudorep1 AND pooledpseudorep2)
out, err = common.run_pipe([
'intersectBed -wa -a %s -b %s %s' %(pooled_peaks_fn, overlap_tr_fn, overlap_pr_fn),
'intersectBed -wa -u -a %s -b stdin' %(pooled_peaks_fn)
], overlapping_peaks_fn)
print "%d peaks overall with true replicates or with pooled pseudorepliates" %(common.count_lines(overlapping_peaks_fn))
'''
#the only difference between the peak_types is how the extra columns are handled
if self.peak_type == "narrowPeak":
awk_command = r"""awk 'BEGIN{FS="\t";OFS="\t"}{s1=$3-$2; s2=$13-$12; if (($21/s1 >= 0.5) || ($21/s2 >= 0.5)) {print $0}}'"""
cut_command = 'cut -f 1-10'
bed_type = 'bed6+4'
elif self.peak_type == "gappedPeak":
awk_command = r"""awk 'BEGIN{FS="\t";OFS="\t"}{s1=$3-$2; s2=$18-$17; if (($31/s1 >= 0.5) || ($31/s2 >= 0.5)) {print $0}}'"""
cut_command = 'cut -f 1-15'
bed_type = 'bed12+3'
elif self.peak_type == "broadPeak":
awk_command = r"""awk 'BEGIN{FS="\t";OFS="\t"}{s1=$3-$2; s2=$12-$11; if (($19/s1 >= 0.5) || ($19/s2 >= 0.5)) {print $0}}'"""
cut_command = 'cut -f 1-9'
bed_type = 'bed6+3'
else:
print "%s is unrecognized. peak_type should be narrowPeak, gappedPeak or broadPeak."
sys.exit()
# Find pooled peaks that overlap Rep1 and Rep2 where overlap is defined 1bp
out, err = common.run_pipe([
'intersectBed -wo -a %s -b %s' %(self.pooled_peaks_fn, self.rep1_peaks_fn),
awk_command,
cut_command,
'sort -u',
'intersectBed -wo -a stdin -b %s' %(self.rep2_peaks_fn),
awk_command,
cut_command,
'sort -u'
], self.overlap_tr_fn)
print "%d peaks overlap with both true replicates" %(common.count_lines(self.overlap_tr_fn))
# Find pooled peaks that overlap PseudoRep1 and PseudoRep2 where overlap is defined as 1bp
out, err = common.run_pipe([
'intersectBed -wo -a %s -b %s' %(self.pooled_peaks_fn, self.pooledpr1_peaks_fn),
awk_command,
cut_command,
'sort -u',
'intersectBed -wo -a stdin -b %s' %(self.pooledpr2_peaks_fn),
awk_command,
cut_command,
'sort -u'
], self.overlap_pr_fn)
print "%d peaks overlap with both pooled pseudoreplicates" %(common.count_lines(self.overlap_pr_fn))
# Combine peak lists
out, err = common.run_pipe([
'cat %s %s' %(self.overlap_tr_fn, self.overlap_pr_fn),
'sort -u'
], self.overlapping_peaks_fn)
print "%d peaks overlap with true replicates or with pooled pseudorepliates" %(common.count_lines(self.overlapping_peaks_fn))
#rejected peaks
out, err = common.run_pipe([
'intersectBed -wa -v -a %s -b %s' %(self.pooled_peaks_fn, self.overlapping_peaks_fn)
], self.rejected_peaks_fn)
print "%d peaks were rejected" %(common.count_lines(self.rejected_peaks_fn))
self.npeaks_in = common.count_lines(common.uncompress(self.pooled_peaks_fn))
self.npeaks_out = common.count_lines(self.overlapping_peaks_fn)
self.npeaks_rejected = common.count_lines(self.rejected_peaks_fn)
#make bigBed files for visualization
self.overlapping_peaks_bb_fn = common.bed2bb(self.overlapping_peaks_fn, self.chrom_sizes_fn, self.as_file_fn, bed_type=bed_type)
self.rejected_peaks_bb_fn = common.bed2bb(self.rejected_peaks_fn, self.chrom_sizes_fn, self.as_file_fn, bed_type=bed_type)
def main(rep1_ta, rep2_ta, ctl1_ta, ctl2_ta, rep1_xcor, rep2_xcor,
npeaks, nodups, rep1_paired_end, rep2_paired_end, chrom_sizes,
spp_version, as_file=None, idr_peaks=False):
assert rep1_paired_end == rep2_paired_end, 'Mixed PE/SE not supported (yet)'
paired_end = rep1_paired_end
rep1_ta_file = dxpy.DXFile(rep1_ta)
rep2_ta_file = dxpy.DXFile(rep2_ta)
unary_control = ctl1_ta == ctl2_ta
ctl1_ta_file = dxpy.DXFile(ctl1_ta)
ctl2_ta_file = dxpy.DXFile(ctl2_ta)
rep1_xcor_file = dxpy.DXFile(rep1_xcor)
rep2_xcor_file = dxpy.DXFile(rep2_xcor)
dxpy.download_dxfile(rep1_ta_file.get_id(), rep1_ta_file.name)
dxpy.download_dxfile(rep2_ta_file.get_id(), rep2_ta_file.name)
dxpy.download_dxfile(ctl1_ta_file.get_id(), ctl1_ta_file.name)
dxpy.download_dxfile(ctl2_ta_file.get_id(), ctl2_ta_file.name)
dxpy.download_dxfile(rep1_xcor_file.get_id(), rep1_xcor_file.name)
dxpy.download_dxfile(rep2_xcor_file.get_id(), rep2_xcor_file.name)
rep1_ta_filename = rep1_ta_file.name
rep2_ta_filename = rep2_ta_file.name
ctl1_ta_filename = ctl1_ta_file.name
ctl2_ta_filename = ctl2_ta_file.name
ntags_rep1 = common.count_lines(rep1_ta_filename)
ntags_rep2 = common.count_lines(rep2_ta_filename)
ntags_ctl1 = common.count_lines(ctl1_ta_filename)
ntags_ctl2 = common.count_lines(ctl2_ta_filename)
for n, name, filename in [
(ntags_rep1, 'replicate 1', rep1_ta_filename),
(ntags_rep2, 'replicate 2', rep2_ta_filename),
(ntags_ctl1, 'control 1', ctl1_ta_filename),
(ntags_ctl2, 'control 2', ctl2_ta_filename)]:
logger.info("Found %d tags in %s file %s" % (n, name, filename))
subprocess.check_call('ls -l', shell=True)
pool_applet = dxpy.find_one_data_object(
classname='applet',
name='pool',
project=dxpy.PROJECT_CONTEXT_ID,
zero_ok=False,
more_ok=False,
return_handler=True)
pool_replicates_subjob = \
pool_applet.run(
{"inputs": [rep1_ta, rep2_ta],
"prefix": 'pooled_reps'},
name='Pool replicates')
pooled_replicates = pool_replicates_subjob.get_output_ref("pooled")
pooled_replicates_xcor_subjob = \
xcor_only(
pooled_replicates,
paired_end,
spp_version,
name='Pool cross-correlation')
rep1_control = ctl1_ta # default. May be changed later.
rep1_ctl_msg = "control rep1"
rep2_control = ctl2_ta # default. May be changed later.
rep2_ctl_msg = "control rep2"
if unary_control:
logger.info("Only one control supplied. Using it for both replicate 1 and 2 and for the pool.")
control_for_pool = rep1_control
pool_ctl_msg = "one control"
else:
pool_controls_subjob = \
pool_applet.run(
{"inputs": [ctl1_ta, ctl2_ta],
"prefix": 'pooled_ctls'},
name='Pool controls')
pooled_controls = pool_controls_subjob.get_output_ref("pooled")
# always use the pooled controls for the pool
control_for_pool = pooled_controls
pool_ctl_msg = "pooled controls"
# use the pooled controls for the reps depending on the ratio of
# rep to control reads
ratio_ctl_reads = float(ntags_ctl1)/float(ntags_ctl2)
if ratio_ctl_reads < 1:
ratio_ctl_reads = 1/ratio_ctl_reads
ratio_cutoff = 1.2
if ratio_ctl_reads > ratio_cutoff:
logger.info(
"Number of reads in controls differ by > factor of %f. Using pooled controls."
% (ratio_cutoff))
rep1_control = pooled_controls
rep1_ctl_msg = "pooled controls"
rep2_control = pooled_controls
rep2_ctl_msg = "pooled controls"
else:
if ntags_ctl1 < ntags_rep1:
logger.info("Fewer reads in control replicate 1 than experiment replicate 1. Using pooled controls for replicate 1.")
rep1_control = pooled_controls
rep1_ctl_msg = "pooled controls"
elif ntags_ctl2 < ntags_rep2:
logger.info("Fewer reads in control replicate 2 than experiment replicate 2. Using pooled controls for replicate 2.")
rep2_control = pooled_controls
rep2_ctl_msg = "pooled controls"
else:
logger.info("Using distinct controls for replicate 1 and 2.")
rep1_ctl_msg = "control rep1"
rep2_ctl_msg = "control rep2"
rep1_peaks_subjob = spp(
rep1_ta,
rep1_control,
rep1_xcor,
chrom_sizes,
spp_version,
bigbed=True,
as_file=as_file,
name='Rep1 peaks vs %s' % (rep1_ctl_msg),
prefix='R1')
rep2_peaks_subjob = spp(
rep2_ta,
rep2_control,
rep2_xcor,
chrom_sizes,
spp_version,
bigbed=True,
as_file=as_file,
name='Rep2 peaks vs %s' % (rep2_ctl_msg),
prefix='R2')
pooled_peaks_subjob = spp(
pooled_replicates,
control_for_pool,
pooled_replicates_xcor_subjob.get_output_ref("CC_scores_file"),
chrom_sizes,
spp_version,
bigbed=True,
as_file=as_file,
name='Pooled peaks vs %s' % (pool_ctl_msg),
prefix='PL')
output = {
'rep1_peaks': rep1_peaks_subjob.get_output_ref("peaks"),
'rep1_peaks_bb': rep1_peaks_subjob.get_output_ref("peaks_bb"),
'rep1_xcor_plot': rep1_peaks_subjob.get_output_ref("xcor_plot"),
'rep1_xcor_scores': rep1_peaks_subjob.get_output_ref("xcor_scores"),
'rep2_peaks': rep2_peaks_subjob.get_output_ref("peaks"),
'rep2_peaks_bb': rep2_peaks_subjob.get_output_ref("peaks_bb"),
'rep2_xcor_plot': rep2_peaks_subjob.get_output_ref("xcor_plot"),
'rep2_xcor_scores': rep2_peaks_subjob.get_output_ref("xcor_scores"),
'pooled_peaks': pooled_peaks_subjob.get_output_ref("peaks"),
'pooled_peaks_bb': pooled_peaks_subjob.get_output_ref("peaks_bb"),
'pooled_xcor_plot': pooled_peaks_subjob.get_output_ref("xcor_plot"),
'pooled_xcor_scores': pooled_peaks_subjob.get_output_ref("xcor_scores")
}
if idr_peaks: # also call peaks on pseudoreplicates for IDR
pseudoreplicator_applet = \
dxpy.find_one_data_object(
classname='applet',
name='pseudoreplicator',
project=dxpy.PROJECT_CONTEXT_ID,
zero_ok=False,
more_ok=False,
return_handler=True)
rep1_pr_subjob = \
pseudoreplicator_applet.run(
{"input_tags": rep1_ta,
"prefix": 'R1PR'},
name='Pseudoreplicate rep1 -> R1PR1,2')
rep2_pr_subjob = \
pseudoreplicator_applet.run(
{"input_tags": rep2_ta,
"prefix": 'R2PR'},
name='Pseudoreplicate rep2 -> R2PR1,2')
pool_pr1_subjob = pool_applet.run({
"inputs": [
rep1_pr_subjob.get_output_ref("pseudoreplicate1"),
rep2_pr_subjob.get_output_ref("pseudoreplicate1")],
"prefix": 'PPR1'},
name='Pool R1PR1+R2PR1 -> PPR1')
pool_pr2_subjob = pool_applet.run({
"inputs": [
rep1_pr_subjob.get_output_ref("pseudoreplicate2"),
rep2_pr_subjob.get_output_ref("pseudoreplicate2")],
"prefix": 'PPR2'},
name='Pool R1PR2+R2PR2 -> PPR2')
# rep1_pr1_xcor_subjob = \
# xcor_only(
# rep1_pr_subjob.get_output_ref("pseudoreplicate1"),
# paired_end,
# spp_version,
# name='R1PR1 cross-correlation')
# rep1_pr2_xcor_subjob = \
# xcor_only(
# rep1_pr_subjob.get_output_ref("pseudoreplicate2"),
# paired_end,
# spp_version,
# name='R1PR2 cross-correlation')
# rep2_pr1_xcor_subjob = \
# xcor_only(
# rep2_pr_subjob.get_output_ref("pseudoreplicate1"),
# paired_end,
# spp_version,
# name='R2PR1 cross-correlation')
# rep2_pr2_xcor_subjob = \
# xcor_only(
# rep2_pr_subjob.get_output_ref("pseudoreplicate2"),
# paired_end,
# spp_version,
# name='R2PR2 cross-correlation')
# pool_pr1_xcor_subjob = \
# xcor_only(
# pool_pr1_subjob.get_output_ref("pooled"),
# paired_end,
# spp_version,
# name='PPR1 cross-correlation')
# pool_pr2_xcor_subjob = \
# xcor_only(
# pool_pr2_subjob.get_output_ref("pooled"),
# paired_end,
# spp_version,
# name='PPR2 cross-correlation')
rep1pr1_peaks_subjob = spp(
rep1_pr_subjob.get_output_ref("pseudoreplicate1"),
rep1_control,
rep1_xcor,
chrom_sizes,
spp_version,
bigbed=False,
name='R1PR1 peaks vs %s' % (rep1_ctl_msg),
prefix='R1PR1')
rep1pr2_peaks_subjob = spp(
rep1_pr_subjob.get_output_ref("pseudoreplicate2"),
rep1_control,
rep1_xcor,
chrom_sizes,
spp_version,
bigbed=False,
name='R1PR2 peaks vs %s' % (rep1_ctl_msg),
prefix='R1PR2')
rep2pr1_peaks_subjob = spp(
rep2_pr_subjob.get_output_ref("pseudoreplicate1"),
rep2_control,
rep2_xcor,
chrom_sizes,
spp_version,
bigbed=False,
name='R2PR1 peaks vs %s' % (rep2_ctl_msg),
prefix='R2PR1')
rep2pr2_peaks_subjob = spp(
rep2_pr_subjob.get_output_ref("pseudoreplicate2"),
rep2_control,
rep2_xcor,
chrom_sizes,
spp_version,
bigbed=False,
name='R2PR2 peaks vs %s' % (rep2_ctl_msg),
prefix='R2PR2')
pooledpr1_peaks_subjob = spp(
pool_pr1_subjob.get_output_ref("pooled"),
control_for_pool,
pooled_replicates_xcor_subjob.get_output_ref("CC_scores_file"),
chrom_sizes,
spp_version,
bigbed=False,
name='PPR1 peaks vs %s' % (pool_ctl_msg),
prefix='PPR1')
pooledpr2_peaks_subjob = spp(
pool_pr2_subjob.get_output_ref("pooled"),
control_for_pool,
pooled_replicates_xcor_subjob.get_output_ref("CC_scores_file"),
chrom_sizes,
spp_version,
bigbed=False,
name='PPR2 peaks vs %s' % (pool_ctl_msg),
prefix='PPR2')
output.update({
'rep1pr1_peaks': rep1pr1_peaks_subjob.get_output_ref("peaks"),
# 'rep1pr1_xcor_plot': rep1pr1_peaks_subjob.get_output_ref("xcor_plot"),
# 'rep1pr1_xcor_scores': rep1pr1_peaks_subjob.get_output_ref("xcor_scores"),
'rep1pr2_peaks': rep1pr2_peaks_subjob.get_output_ref("peaks"),
# 'rep1pr2_xcor_plot': rep1pr2_peaks_subjob.get_output_ref("xcor_plot"),
# 'rep1pr2_xcor_scores': rep1pr2_peaks_subjob.get_output_ref("xcor_scores"),
'rep2pr1_peaks': rep2pr1_peaks_subjob.get_output_ref("peaks"),
# 'rep2pr1_xcor_plot': rep2pr1_peaks_subjob.get_output_ref("xcor_plot"),
# 'rep2pr1_xcor_scores': rep2pr1_peaks_subjob.get_output_ref("xcor_scores"),
'rep2pr2_peaks': rep2pr2_peaks_subjob.get_output_ref("peaks"),
# 'rep2pr2_xcor_plot': rep2pr2_peaks_subjob.get_output_ref("xcor_plot"),
# 'rep2pr2_xcor_scores': rep2pr2_peaks_subjob.get_output_ref("xcor_scores"),
'pooledpr1_peaks': pooledpr1_peaks_subjob.get_output_ref("peaks"),
# 'pooledpr1_xcor_plot': pooledpr1_peaks_subjob.get_output_ref("xcor_plot"),
# 'pooledpr1_xcor_scores': pooledpr1_peaks_subjob.get_output_ref("xcor_scores"),
'pooledpr2_peaks': pooledpr2_peaks_subjob.get_output_ref("peaks"),
# 'pooledpr2_xcor_plot': pooledpr2_peaks_subjob.get_output_ref("xcor_plot"),
# 'pooledpr2_xcor_scores': pooledpr2_peaks_subjob.get_output_ref("xcor_scores"),
})
return output
