def fast_rnaseq(samples, run_parallel):
to_index = determine_indexes_to_make(samples)
run_parallel("run_salmon_index", [to_index])
samples = run_parallel("run_salmon_reads", samples)
samples = run_parallel("run_counts_spikein", samples)
samples = spikein.combine_spikein(samples)
return samples
def run_cluster(*data):
"""
Run seqcluster cluster to detect smallRNA clusters
"""
sample = data[0][0]
tools = dd.get_expression_caller(data[0][0])
work_dir = dd.get_work_dir(sample)
out_dir = op.join(work_dir, "seqcluster", "cluster")
out_dir = op.abspath(safe_makedir(out_dir))
prepare_dir = op.join(work_dir, "seqcluster", "prepare")
bam_file = data[0][0]["cluster_bam"]
if "seqcluster" in tools:
gtf_file = dd.get_transcriptome_gtf(sample) if dd.get_transcriptome_gtf(sample) else dd.get_srna_gtf_file(sample)
sample["seqcluster"] = _cluster(bam_file, data[0][0]["seqcluster_prepare_ma"],
out_dir, dd.get_ref_file(sample),
gtf_file)
sample["report"] = _report(sample, dd.get_ref_file(sample))
if "mirge" in tools:
sample["mirge"] = mirge.run(data)
out_mirna = _make_isomir_counts(data, out_dir=op.join(work_dir, "mirbase"))
if out_mirna:
sample = dd.set_mirna_counts(sample, out_mirna[0])
sample = dd.set_isomir_counts(sample, out_mirna[1])
out_novel = _make_isomir_counts(data, "seqbuster_novel", op.join(work_dir, "mirdeep2"), "_novel")
if out_novel:
sample = dd.set_novel_mirna_counts(sample, out_novel[0])
sample = dd.set_novel_isomir_counts(sample, out_novel[1])
data[0][0] = sample
data = spikein.combine_spikein(data)
return data
def run_cluster(*data):
"""
Run seqcluster cluster to detect smallRNA clusters
"""
sample = data[0][0]
tools = dd.get_expression_caller(data[0][0])
work_dir = dd.get_work_dir(sample)
out_dir = op.join(work_dir, "seqcluster", "cluster")
out_dir = op.abspath(safe_makedir(out_dir))
prepare_dir = op.join(work_dir, "seqcluster", "prepare")
bam_file = data[0][0]["work_bam"]
if "seqcluster" in tools:
sample["seqcluster"] = _cluster(bam_file, data[0][0]["seqcluster_prepare_ma"], out_dir, dd.get_ref_file(sample), dd.get_srna_gtf_file(sample))
sample["report"] = _report(sample, dd.get_ref_file(sample))
out_mirna = _make_isomir_counts(data, out_dir=op.join(work_dir, "mirbase"))
if out_mirna:
sample = dd.set_mirna_counts(sample, out_mirna[0])
sample = dd.set_isomir_counts(sample, out_mirna[1])
out_novel = _make_isomir_counts(data, "seqbuster_novel", op.join(work_dir, "mirdeep2"), "_novel")
if out_novel:
sample = dd.set_novel_mirna_counts(sample, out_novel[0])
sample = dd.set_novel_isomir_counts(sample, out_novel[1])
data[0][0] = sample
data = spikein.combine_spikein(data)
return data
def combine_files(samples):
"""
after quantitation, combine the counts/FPKM/TPM/etc into a single table with
all samples
"""
gtf_file = dd.get_gtf_file(samples[0][0], None)
dexseq_gff = dd.get_dexseq_gff(samples[0][0])
# combine featureCount files
count_files = filter_missing([dd.get_count_file(x[0]) for x in samples])
combined = count.combine_count_files(count_files, ext=".counts")
annotated = count.annotate_combined_count_file(combined, gtf_file)
# combine eXpress files
express_counts_combined = combine_express(samples, combined)
# combine Cufflinks files
fpkm_combined_file = os.path.splitext(combined)[0] + ".fpkm"
fpkm_files = filter_missing([dd.get_fpkm(x[0]) for x in samples])
if fpkm_files:
fpkm_combined = count.combine_count_files(fpkm_files, fpkm_combined_file)
else:
fpkm_combined = None
fpkm_isoform_combined_file = os.path.splitext(combined)[0] + ".isoform.fpkm"
isoform_files = filter_missing([dd.get_fpkm_isoform(x[0]) for x in samples])
if isoform_files:
fpkm_isoform_combined = count.combine_count_files(isoform_files,
fpkm_isoform_combined_file,
".isoform.fpkm")
else:
fpkm_isoform_combined = None
# combine DEXseq files
dexseq_combined_file = os.path.splitext(combined)[0] + ".dexseq"
to_combine_dexseq = filter_missing([dd.get_dexseq_counts(data[0]) for data in samples])
if to_combine_dexseq:
dexseq_combined = count.combine_count_files(to_combine_dexseq,
dexseq_combined_file, ".dexseq")
dexseq.create_dexseq_annotation(dexseq_gff, dexseq_combined)
else:
dexseq_combined = None
samples = spikein.combine_spikein(samples)
updated_samples = []
for data in dd.sample_data_iterator(samples):
data = dd.set_combined_counts(data, combined)
if annotated:
data = dd.set_annotated_combined_counts(data, annotated)
if fpkm_combined:
data = dd.set_combined_fpkm(data, fpkm_combined)
if fpkm_isoform_combined:
data = dd.set_combined_fpkm_isoform(data, fpkm_isoform_combined)
if express_counts_combined:
data = dd.set_express_counts(data, express_counts_combined['counts'])
data = dd.set_express_tpm(data, express_counts_combined['tpm'])
data = dd.set_express_fpkm(data, express_counts_combined['fpkm'])
data = dd.set_isoform_to_gene(data, express_counts_combined['isoform_to_gene'])
if dexseq_combined:
data = dd.set_dexseq_counts(data, dexseq_combined_file)
updated_samples.append([data])
return updated_samples
def combine_files(samples):
"""
after quantitation, combine the counts/FPKM/TPM/etc into a single table with
all samples
"""
gtf_file = dd.get_gtf_file(samples[0][0], None)
dexseq_gff = dd.get_dexseq_gff(samples[0][0])
# combine featureCount files
count_files = filter_missing([dd.get_count_file(x[0]) for x in samples])
combined = count.combine_count_files(count_files, ext=".counts")
annotated = count.annotate_combined_count_file(combined, gtf_file)
# combine eXpress files
express_counts_combined = combine_express(samples, combined)
# combine Cufflinks files
fpkm_combined_file = os.path.splitext(combined)[0] + ".fpkm"
fpkm_files = filter_missing([dd.get_fpkm(x[0]) for x in samples])
if fpkm_files:
fpkm_combined = count.combine_count_files(fpkm_files, fpkm_combined_file)
else:
fpkm_combined = None
fpkm_isoform_combined_file = os.path.splitext(combined)[0] + ".isoform.fpkm"
isoform_files = filter_missing([dd.get_fpkm_isoform(x[0]) for x in samples])
if isoform_files:
fpkm_isoform_combined = count.combine_count_files(isoform_files,
fpkm_isoform_combined_file,
".isoform.fpkm")
else:
fpkm_isoform_combined = None
# combine DEXseq files
dexseq_combined_file = os.path.splitext(combined)[0] + ".dexseq"
to_combine_dexseq = filter_missing([dd.get_dexseq_counts(data[0]) for data in samples])
if to_combine_dexseq:
dexseq_combined = count.combine_count_files(to_combine_dexseq,
dexseq_combined_file, ".dexseq")
dexseq.create_dexseq_annotation(dexseq_gff, dexseq_combined)
else:
dexseq_combined = None
samples = spikein.combine_spikein(samples)
updated_samples = []
for data in dd.sample_data_iterator(samples):
data = dd.set_combined_counts(data, combined)
if annotated:
data = dd.set_annotated_combined_counts(data, annotated)
if fpkm_combined:
data = dd.set_combined_fpkm(data, fpkm_combined)
if fpkm_isoform_combined:
data = dd.set_combined_fpkm_isoform(data, fpkm_isoform_combined)
if express_counts_combined:
data = dd.set_express_counts(data, express_counts_combined['counts'])
data = dd.set_express_tpm(data, express_counts_combined['tpm'])
data = dd.set_express_fpkm(data, express_counts_combined['fpkm'])
data = dd.set_isoform_to_gene(data, express_counts_combined['isoform_to_gene'])
if dexseq_combined:
data = dd.set_dexseq_counts(data, dexseq_combined_file)
updated_samples.append([data])
return updated_samples
def combine_files(samples):
"""
after quantitation, combine the counts/FPKM/TPM/etc into a single table with
all samples
"""
data = samples[0][0]
# prefer the supplied transcriptome gtf file
gtf_file = dd.get_transcriptome_gtf(data, None)
if not gtf_file:
gtf_file = dd.get_gtf_file(data, None)
dexseq_gff = dd.get_dexseq_gff(data)
# combine featureCount files
count_files = filter_missing([dd.get_count_file(x[0]) for x in samples])
combined = count.combine_count_files(count_files, ext=".counts")
annotated = count.annotate_combined_count_file(combined, gtf_file)
# add tx2gene file
tx2gene_file = os.path.join(dd.get_work_dir(data), "annotation",
"tx2gene.csv")
if gtf_file:
tx2gene_file = sailfish.create_combined_tx2gene(data)
# combine eXpress files
express_counts_combined = combine_express(samples, combined)
# combine Cufflinks files
fpkm_files = filter_missing([dd.get_fpkm(x[0]) for x in samples])
if fpkm_files:
fpkm_combined_file = os.path.splitext(combined)[0] + ".fpkm"
fpkm_combined = count.combine_count_files(fpkm_files,
fpkm_combined_file)
else:
fpkm_combined = None
isoform_files = filter_missing(
[dd.get_fpkm_isoform(x[0]) for x in samples])
if isoform_files:
fpkm_isoform_combined_file = os.path.splitext(
combined)[0] + ".isoform.fpkm"
fpkm_isoform_combined = count.combine_count_files(
isoform_files, fpkm_isoform_combined_file, ".isoform.fpkm")
else:
fpkm_isoform_combined = None
# combine DEXseq files
to_combine_dexseq = filter_missing(
[dd.get_dexseq_counts(data[0]) for data in samples])
if to_combine_dexseq:
dexseq_combined_file = os.path.splitext(combined)[0] + ".dexseq"
dexseq_combined = count.combine_count_files(to_combine_dexseq,
dexseq_combined_file,
".dexseq")
dexseq.create_dexseq_annotation(dexseq_gff, dexseq_combined)
else:
dexseq_combined = None
samples = spikein.combine_spikein(samples)
updated_samples = []
for data in dd.sample_data_iterator(samples):
if combined:
data = dd.set_combined_counts(data, combined)
if annotated:
data = dd.set_annotated_combined_counts(data, annotated)
if fpkm_combined:
data = dd.set_combined_fpkm(data, fpkm_combined)
if fpkm_isoform_combined:
data = dd.set_combined_fpkm_isoform(data, fpkm_isoform_combined)
if express_counts_combined:
data = dd.set_express_counts(data,
express_counts_combined['counts'])
data = dd.set_express_tpm(data, express_counts_combined['tpm'])
data = dd.set_express_fpkm(data, express_counts_combined['fpkm'])
data = dd.set_isoform_to_gene(
data, express_counts_combined['isoform_to_gene'])
if dexseq_combined:
data = dd.set_dexseq_counts(data, dexseq_combined_file)
if gtf_file:
data = dd.set_tx2gene(data, tx2gene_file)
updated_samples.append([data])
return updated_samples
def fast_rnaseq(samples, run_parallel):
samples = run_parallel("run_salmon_index", [samples])
samples = run_parallel("run_salmon_reads", samples)
samples = run_parallel("run_counts_spikein", samples)
samples = spikein.combine_spikein(samples)
return samples
def combine_files(samples):
"""
after quantitation, combine the counts/FPKM/TPM/etc into a single table with
all samples
"""
data = samples[0][0]
# prefer the supplied transcriptome gtf file
gtf_file = dd.get_transcriptome_gtf(data, None)
if not gtf_file:
gtf_file = dd.get_gtf_file(data, None)
dexseq_gff = dd.get_dexseq_gff(data)
# combine featureCount files
count_files = filter_missing([dd.get_count_file(x[0]) for x in samples])
combined = count.combine_count_files(count_files, ext=".counts")
annotated = count.annotate_combined_count_file(combined, gtf_file)
# add tx2gene file
tx2gene_file = os.path.join(dd.get_work_dir(data), "annotation", "tx2gene.csv")
if gtf_file:
tx2gene_file = sailfish.create_combined_tx2gene(data)
# combine eXpress files
express_counts_combined = combine_express(samples, combined)
# combine Cufflinks files
fpkm_files = filter_missing([dd.get_fpkm(x[0]) for x in samples])
if fpkm_files and combined:
fpkm_combined_file = os.path.splitext(combined)[0] + ".fpkm"
fpkm_combined = count.combine_count_files(fpkm_files, fpkm_combined_file)
else:
fpkm_combined = None
isoform_files = filter_missing([dd.get_fpkm_isoform(x[0]) for x in samples])
if isoform_files and combined:
fpkm_isoform_combined_file = os.path.splitext(combined)[0] + ".isoform.fpkm"
fpkm_isoform_combined = count.combine_count_files(isoform_files,
fpkm_isoform_combined_file,
".isoform.fpkm")
else:
fpkm_isoform_combined = None
# combine DEXseq files
to_combine_dexseq = filter_missing([dd.get_dexseq_counts(data[0]) for data
in samples])
if to_combine_dexseq and combined:
dexseq_combined_file = os.path.splitext(combined)[0] + ".dexseq"
dexseq_combined = count.combine_count_files(to_combine_dexseq,
dexseq_combined_file, ".dexseq")
if dexseq_combined:
dexseq.create_dexseq_annotation(dexseq_gff, dexseq_combined)
else:
dexseq_combined = None
samples = spikein.combine_spikein(samples)
updated_samples = []
for data in dd.sample_data_iterator(samples):
if combined:
data = dd.set_combined_counts(data, combined)
if annotated:
data = dd.set_annotated_combined_counts(data, annotated)
if fpkm_combined:
data = dd.set_combined_fpkm(data, fpkm_combined)
if fpkm_isoform_combined:
data = dd.set_combined_fpkm_isoform(data, fpkm_isoform_combined)
if express_counts_combined:
data = dd.set_express_counts(data, express_counts_combined['counts'])
data = dd.set_express_tpm(data, express_counts_combined['tpm'])
data = dd.set_express_fpkm(data, express_counts_combined['fpkm'])
data = dd.set_isoform_to_gene(data, express_counts_combined['isoform_to_gene'])
if dexseq_combined:
data = dd.set_dexseq_counts(data, dexseq_combined_file)
if gtf_file:
data = dd.set_tx2gene(data, tx2gene_file)
updated_samples.append([data])
return updated_samples
def fast_rnaseq(samples, run_parallel):
samples = run_parallel("run_salmon_index", [samples])
samples = run_parallel("run_salmon_reads", samples)
samples = run_parallel("run_counts_spikein", samples)
samples = spikein.combine_spikein(samples)
return samples