def run(data):
"""Quantitaive isoforms expression by eXpress"""
name = dd.get_sample_name(data)
in_bam = dd.get_transcriptome_bam(data)
config = data['config']
if not in_bam:
logger.info("Transcriptome-mapped BAM file not found, skipping eXpress.")
return data
gtf_fasta = gtf.gtf_to_fasta(dd.get_gtf_file(data), dd.get_ref_file(data))
out_dir = os.path.join(dd.get_work_dir(data), "express", name)
out_file = os.path.join(out_dir, name + ".xprs")
express = config_utils.get_program("express", data['config'])
strand = _set_stranded_flag(in_bam, data)
if not file_exists(out_file):
with tx_tmpdir(data) as tmp_dir:
with file_transaction(out_dir) as tx_out_dir:
bam_file = _prepare_bam_file(in_bam, tmp_dir, config)
cmd = ("{express} --no-update-check -o {tx_out_dir} {strand} {gtf_fasta} {bam_file}")
do.run(cmd.format(**locals()), "Run express on %s." % in_bam, {})
shutil.move(os.path.join(out_dir, "results.xprs"), out_file)
eff_count_file = _get_column(out_file, out_file.replace(".xprs", "_eff.counts"), 7)
tpm_file = _get_column(out_file, out_file.replace("xprs", "tpm"), 14)
fpkm_file = _get_column(out_file, out_file.replace("xprs", "fpkm"), 10)
data = dd.set_express_counts(data, eff_count_file)
data = dd.set_express_tpm(data, tpm_file)
data = dd.set_express_fpkm(data, fpkm_file)
return data
def run(data):
"""Quantitaive isoforms expression by eXpress"""
name = dd.get_sample_name(data)
in_bam = dd.get_transcriptome_bam(data)
if not in_bam:
logger.info(
"Transcriptome-mapped BAM file not found, skipping eXpress.")
return data
gtf_fasta = gtf.gtf_to_fasta(dd.get_gtf_file(data), dd.get_ref_file(data))
out_dir = os.path.join(dd.get_work_dir(data), "express", name)
out_file = os.path.join(out_dir, name + ".xprs")
express = config_utils.get_program("express", data['config'])
strand = _set_stranded_flag(in_bam, data)
if not file_exists(out_file):
with file_transaction(out_dir) as tx_out_dir:
cmd = (
"{express} --no-update-check -o {tx_out_dir} {strand} {gtf_fasta} {in_bam}"
)
do.run(cmd.format(**locals()), "Run express on %s." % in_bam, {})
shutil.move(os.path.join(out_dir, "results.xprs"), out_file)
eff_count_file = _get_column(out_file,
out_file.replace(".xprs", "_eff.counts"), 7)
tpm_file = _get_column(out_file, out_file.replace("xprs", "tpm"), 14)
fpkm_file = _get_column(out_file, out_file.replace("xprs", "fpkm"), 10)
data = dd.set_express_counts(data, eff_count_file)
data = dd.set_express_tpm(data, tpm_file)
data = dd.set_express_fpkm(data, fpkm_file)
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 estimate_expression(samples, run_parallel):
samples = run_parallel("generate_transcript_counts", samples)
count_files = filter_missing([dd.get_count_file(x[0]) for x in samples])
combined = count.combine_count_files(count_files)
gtf_file = dd.get_gtf_file(samples[0][0], None)
annotated = count.annotate_combined_count_file(combined, gtf_file)
samples = run_parallel("run_express", samples)
express_counts_combined = combine_express(samples, combined)
samples = run_parallel("run_cufflinks", samples)
#gene
fpkm_combined_file = os.path.splitext(combined)[0] + ".fpkm"
fpkm_files = filter_missing([dd.get_fpkm(x[0]) for x in samples])
fpkm_combined = count.combine_count_files(fpkm_files, fpkm_combined_file)
#isoform
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])
fpkm_isoform_combined = count.combine_count_files(isoform_files,
fpkm_isoform_combined_file,
".isoform.fpkm")
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")
else:
dexseq_combined = None
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_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'])
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 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