def index(in_bam, config):
"""Index a BAM file, skipping if index present.
Centralizes BAM indexing providing ability to switch indexing approaches.
"""
assert is_bam(in_bam), "%s in not a BAM file" % in_bam
index_file = "%s.bai" % in_bam
alt_index_file = "%s.bai" % os.path.splitext(in_bam)[0]
if (not utils.file_uptodate(index_file, in_bam)
and not utils.file_uptodate(alt_index_file, in_bam)):
# Remove old index files and re-run to prevent linking into tx directory
for fname in [index_file, alt_index_file]:
utils.remove_safe(fname)
sambamba = _get_sambamba(config)
samtools = config_utils.get_program("samtools", config)
num_cores = config["algorithm"].get("num_cores", 1)
with file_transaction(config, index_file) as tx_index_file:
assert tx_index_file.find(".bam.bai") > 0
tx_bam_file = tx_index_file.replace(".bam.bai", ".bam")
utils.symlink_plus(in_bam, tx_bam_file)
if sambamba:
cmd = "{sambamba} index -t {num_cores} {tx_bam_file}"
else:
cmd = "{samtools} index {tx_bam_file}"
do.run(cmd.format(**locals()),
"Index BAM file: %s" % os.path.basename(in_bam))
return index_file if utils.file_uptodate(index_file,
in_bam) else alt_index_file
def _run_scalpel_paired(align_bams, items, ref_file, assoc_files,
region=None, out_file=None):
"""Detect indels with Scalpel.
This is used for paired tumor / normal samples.
"""
config = items[0]["config"]
if out_file is None:
out_file = "%s-paired-variants.vcf.gz" % os.path.splitext(align_bams[0])[0]
if not utils.file_exists(out_file):
with file_transaction(config, out_file) as tx_out_file:
paired = get_paired_bams(align_bams, items)
if not paired.normal_bam:
ann_file = _run_scalpel_caller(align_bams, items, ref_file,
assoc_files, region, out_file)
return ann_file
vcfstreamsort = config_utils.get_program("vcfstreamsort", config)
perl_exports = utils.get_perl_exports(os.path.dirname(tx_out_file))
tmp_path = "%s-scalpel-work" % utils.splitext_plus(out_file)[0]
db_file = os.path.join(tmp_path, "main", "somatic.db")
if not os.path.exists(db_file + ".dir"):
if os.path.exists(tmp_path):
utils.remove_safe(tmp_path)
opts = " ".join(_scalpel_options_from_config(items, config, out_file, region, tmp_path))
opts += " --ref {}".format(ref_file)
opts += " --dir %s" % tmp_path
# caling
cl = ("{perl_exports} && "
"scalpel-discovery --somatic {opts} --tumor {paired.tumor_bam} --normal {paired.normal_bam}")
do.run(cl.format(**locals()), "Genotyping paired variants with Scalpel", {})
# filtering to adjust input parameters
bed_opts = " ".join(_scalpel_bed_file_opts(items, config, out_file, region, tmp_path))
use_defaults = True
if use_defaults:
scalpel_tmp_file = os.path.join(tmp_path, "main/somatic.indel.vcf")
# Uses default filters but can tweak min-alt-count-tumor and min-phred-fisher
# to swap precision for sensitivity
else:
scalpel_tmp_file = os.path.join(tmp_path, "main/somatic-indel-filter.vcf.gz")
with file_transaction(config, scalpel_tmp_file) as tx_indel_file:
cmd = ("{perl_exports} && "
"scalpel-export --somatic {bed_opts} --ref {ref_file} --db {db_file} "
"--min-alt-count-tumor 5 --min-phred-fisher 10 --min-vaf-tumor 0.1 "
"| bgzip -c > {tx_indel_file}")
do.run(cmd.format(**locals()), "Scalpel somatic indel filter", {})
scalpel_tmp_file = bgzip_and_index(scalpel_tmp_file, config)
scalpel_tmp_file_common = bgzip_and_index(os.path.join(tmp_path, "main/common.indel.vcf"), config)
compress_cmd = "| bgzip -c" if out_file.endswith("gz") else ""
bcftools_cmd_chi2 = get_scalpel_bcftools_filter_expression("chi2", config)
bcftools_cmd_common = get_scalpel_bcftools_filter_expression("reject", config)
fix_ambig = vcfutils.fix_ambiguous_cl()
cl2 = ("vcfcat <({bcftools_cmd_chi2} {scalpel_tmp_file}) "
"<({bcftools_cmd_common} {scalpel_tmp_file_common}) | "
" {fix_ambig} | {vcfstreamsort} {compress_cmd} > {tx_out_file}")
do.run(cl2.format(**locals()), "Finalising Scalpel variants", {})
ann_file = annotation.annotate_nongatk_vcf(out_file, align_bams,
assoc_files.get("dbsnp"), ref_file,
config)
return ann_file
def index(in_bam, config, check_timestamp=True):
"""Index a BAM file, skipping if index present.
Centralizes BAM indexing providing ability to switch indexing approaches.
"""
assert is_bam(in_bam), "%s in not a BAM file" % in_bam
index_file = "%s.bai" % in_bam
alt_index_file = "%s.bai" % os.path.splitext(in_bam)[0]
if check_timestamp:
bai_exists = utils.file_uptodate(index_file, in_bam) or utils.file_uptodate(alt_index_file, in_bam)
else:
bai_exists = utils.file_exists(index_file) or utils.file_exists(alt_index_file)
if not bai_exists:
# Remove old index files and re-run to prevent linking into tx directory
for fname in [index_file, alt_index_file]:
utils.remove_safe(fname)
sambamba = _get_sambamba(config)
assert sambamba, "Did not find sambamba for indexing"
samtools = config_utils.get_program("samtools", config)
num_cores = config["algorithm"].get("num_cores", 1)
with file_transaction(config, index_file) as tx_index_file:
assert tx_index_file.find(".bam.bai") > 0
tx_bam_file = tx_index_file.replace(".bam.bai", ".bam")
utils.symlink_plus(in_bam, tx_bam_file)
try:
cmd = "{sambamba} index -t {num_cores} {tx_bam_file}"
do.run(cmd.format(**locals()), "Index BAM file with sambamba: %s" % os.path.basename(in_bam))
except subprocess.CalledProcessError:
cmd = "{samtools} index {in_bam} {tx_index_file}"
do.run(cmd.format(**locals()), "Backup single thread index of BAM file with samtools: %s"
% os.path.basename(in_bam))
return index_file if utils.file_exists(index_file) else alt_index_file
def index(in_bam, config):
"""Index a BAM file, skipping if index present.
Centralizes BAM indexing providing ability to switch indexing approaches.
"""
assert is_bam(in_bam), "%s in not a BAM file" % in_bam
index_file = "%s.bai" % in_bam
alt_index_file = "%s.bai" % os.path.splitext(in_bam)[0]
if (not utils.file_uptodate(index_file, in_bam) and
not utils.file_uptodate(alt_index_file, in_bam)):
# Remove old index files and re-run to prevent linking into tx directory
for fname in [index_file, alt_index_file]:
utils.remove_safe(fname)
sambamba = _get_sambamba(config)
samtools = config_utils.get_program("samtools", config)
num_cores = config["algorithm"].get("num_cores", 1)
with file_transaction(config, index_file) as tx_index_file:
assert tx_index_file.find(".bam.bai") > 0
tx_bam_file = tx_index_file.replace(".bam.bai", ".bam")
utils.symlink_plus(in_bam, tx_bam_file)
if sambamba:
cmd = "{sambamba} index -t {num_cores} {tx_bam_file}"
else:
cmd = "{samtools} index {tx_bam_file}"
do.run(cmd.format(**locals()), "Index BAM file: %s" % os.path.basename(in_bam))
return index_file if utils.file_uptodate(index_file, in_bam) else alt_index_file
def _move_file_with_sizecheck(tx_file, final_file):
"""Move transaction file to final location,
with size checks avoiding failed transfers.
Creates an empty file with '.bcbiotmp' extention in the destination
location, which serves as a flag. If a file like that is present,
it means that transaction didn't finish successfully.
"""
logger.debug("Moving %s to %s" % (tx_file, final_file))
tmp_file = final_file + ".bcbiotmp"
open(tmp_file, 'wb').close()
want_size = utils.get_size(tx_file)
shutil.move(tx_file, final_file)
transfer_size = utils.get_size(final_file)
assert want_size == transfer_size, (
'distributed.transaction.file_transaction: File copy error: '
'file or directory on temporary storage ({}) size {} bytes '
'does not equal size of file or directory after transfer to '
'shared storage ({}) size {} bytes'.format(tx_file, want_size,
final_file, transfer_size))
utils.remove_safe(tmp_file)
def _install_kraken_db(datadir, args):
"""Install kraken minimal DB in genome folder.
"""
kraken = os.path.join(datadir, "genomes/kraken")
url = "https://ccb.jhu.edu/software/kraken/dl/minikraken.tgz"
compress = os.path.join(kraken, os.path.basename(url))
base, ext = utils.splitext_plus(os.path.basename(url))
db = os.path.join(kraken, base)
tooldir = args.tooldir or get_defaults()["tooldir"]
if os.path.exists(os.path.join(tooldir, "bin", "kraken")):
if not os.path.exists(kraken):
utils.safe_makedir(kraken)
if not os.path.exists(db):
if not os.path.exists(compress):
subprocess.check_call(
["wget", "-O", compress, url, "--no-check-certificate"])
cmd = ["tar", "-xzvf", compress, "-C", kraken]
subprocess.check_call(cmd)
shutil.move(os.path.join(kraken, "minikraken_20140330"),
os.path.join(kraken, "minikraken"))
utils.remove_safe(compress)
else:
raise argparse.ArgumentTypeError(
"kraken not installed in tooldir %s." %
os.path.join(tooldir, "bin", "kraken"))
def _move_file_with_sizecheck(tx_file, final_file):
"""Move transaction file to final location,
with size checks avoiding failed transfers.
Creates an empty file with '.bcbiotmp' extention in the destination
location, which serves as a flag. If a file like that is present,
it means that transaction didn't finish successfully.
"""
#logger.debug("Moving %s to %s" % (tx_file, final_file))
tmp_file = final_file + ".bcbiotmp"
open(tmp_file, 'wb').close()
want_size = utils.get_size(tx_file)
shutil.move(tx_file, final_file)
transfer_size = utils.get_size(final_file)
assert want_size == transfer_size, (
'distributed.transaction.file_transaction: File copy error: '
'file or directory on temporary storage ({}) size {} bytes '
'does not equal size of file or directory after transfer to '
'shared storage ({}) size {} bytes'.format(
tx_file, want_size, final_file, transfer_size)
)
utils.remove_safe(tmp_file)
def _run_workflow(data, workflow_file, work_dir):
"""Run Strelka2 analysis inside prepared workflow directory.
"""
utils.remove_safe(os.path.join(work_dir, "workspace"))
cmd = [sys.executable, workflow_file, "-m", "local", "-j", dd.get_num_cores(data), "--quiet"]
do.run(cmd, "Run Strelka2: %s" % dd.get_sample_name(data))
utils.remove_safe(os.path.join(work_dir, "workspace"))
def _install_kraken_db(datadir, args):
"""Install kraken minimal DB in genome folder.
"""
kraken = os.path.join(datadir, "genomes/kraken")
url = "https://ccb.jhu.edu/software/kraken/dl/minikraken.tgz"
compress = os.path.join(kraken, os.path.basename(url))
base, ext = utils.splitext_plus(os.path.basename(url))
db = os.path.join(kraken, base)
tooldir = args.tooldir or get_defaults()["tooldir"]
requests.packages.urllib3.disable_warnings()
last_mod = urllib.urlopen(url).info().getheader("Last-Modified")
last_mod = dateutil.parser.parse(last_mod).astimezone(dateutil.tz.tzutc())
if os.path.exists(os.path.join(tooldir, "bin", "kraken")):
if not os.path.exists(db):
is_new_version = True
else:
cur_file = glob.glob(os.path.join(kraken, "minikraken_*"))[0]
cur_version = datetime.datetime.utcfromtimestamp(os.path.getmtime(cur_file))
is_new_version = last_mod.date() > cur_version.date()
if is_new_version:
shutil.move(cur_file, cur_file.replace("minikraken", "old"))
if not os.path.exists(kraken):
utils.safe_makedir(kraken)
if is_new_version:
if not os.path.exists(compress):
subprocess.check_call(["wget", "-O", compress, url, "--no-check-certificate"])
cmd = ["tar", "-xzvf", compress, "-C", kraken]
subprocess.check_call(cmd)
last_version = glob.glob(os.path.join(kraken, "minikraken_*"))
utils.symlink_plus(os.path.join(kraken, last_version[0]), os.path.join(kraken, "minikraken"))
utils.remove_safe(compress)
else:
print "You have the latest version %s." % last_mod
else:
raise argparse.ArgumentTypeError("kraken not installed in tooldir %s." % os.path.join(tooldir, "bin", "kraken"))
def _run_workflow(data, workflow_file, work_dir):
"""Run Strelka2 analysis inside prepared workflow directory.
"""
utils.remove_safe(os.path.join(work_dir, "workspace"))
cmd = [sys.executable, workflow_file, "-m", "local", "-j", dd.get_num_cores(data), "--quiet"]
do.run(cmd, "Run Strelka2: %s" % dd.get_sample_name(data))
utils.remove_safe(os.path.join(work_dir, "workspace"))
def index(in_bam, config, check_timestamp=True):
"""Index a BAM file, skipping if index present.
Centralizes BAM indexing providing ability to switch indexing approaches.
"""
assert is_bam(in_bam), "%s in not a BAM file" % in_bam
index_file = "%s.bai" % in_bam
alt_index_file = "%s.bai" % os.path.splitext(in_bam)[0]
if check_timestamp:
bai_exists = utils.file_uptodate(
index_file, in_bam) or utils.file_uptodate(alt_index_file, in_bam)
else:
bai_exists = utils.file_exists(index_file) or utils.file_exists(
alt_index_file)
if not bai_exists:
# Remove old index files and re-run to prevent linking into tx directory
for fname in [index_file, alt_index_file]:
utils.remove_safe(fname)
samtools = config_utils.get_program("samtools", config)
num_cores = config["algorithm"].get("num_cores", 1)
with file_transaction(config, index_file) as tx_index_file:
cmd = "{samtools} index -@ {num_cores} {in_bam} {tx_index_file}"
do.run(cmd.format(**locals()),
"Index BAM file: %s" % os.path.basename(in_bam))
return index_file if utils.file_exists(index_file) else alt_index_file
def _run_workflow(data, workflow_file, work_dir):
"""Run Strelka2 analysis inside prepared workflow directory.
"""
utils.remove_safe(os.path.join(work_dir, "workspace"))
cmd = [utils.get_program_python("configureStrelkaGermlineWorkflow.py"),
workflow_file, "-m", "local", "-j", dd.get_num_cores(data), "--quiet"]
do.run(cmd, "Run Strelka2: %s" % dd.get_sample_name(data))
utils.remove_safe(os.path.join(work_dir, "workspace"))
def _run_workflow(items, paired, workflow_file, work_dir):
"""Run manta analysis inside prepared workflow directory.
"""
utils.remove_safe(os.path.join(work_dir, "workspace"))
data = paired.tumor_data if paired else items[0]
cmd = [sys.executable, workflow_file, "-m", "local", "-j", dd.get_num_cores(data)]
do.run(cmd, "Run manta SV analysis")
utils.remove_safe(os.path.join(work_dir, "workspace"))
def remove(in_bam):
"""
remove bam file and the index if exists
"""
if utils.file_exists(in_bam):
utils.remove_safe(in_bam)
if utils.file_exists(in_bam + ".bai"):
utils.remove_safe(in_bam + ".bai")
def _run_workflow(items, paired, workflow_file, work_dir):
"""Run manta analysis inside prepared workflow directory.
"""
utils.remove_safe(os.path.join(work_dir, "workspace"))
data = paired.tumor_data if paired else items[0]
cmd = [sys.executable, workflow_file, "-m", "local", "-j", dd.get_num_cores(data)]
do.run(cmd, "Run manta SV analysis")
utils.remove_safe(os.path.join(work_dir, "workspace"))
def remove(in_bam):
"""
remove bam file and the index if exists
"""
if utils.file_exists(in_bam):
utils.remove_safe(in_bam)
if utils.file_exists(in_bam + ".bai"):
utils.remove_safe(in_bam + ".bai")
def _run_scalpel_caller(align_bams,
items,
ref_file,
assoc_files,
region=None,
out_file=None):
"""Detect indels with Scalpel.
Single sample mode.
"""
config = items[0]["config"]
if out_file is None:
out_file = "%s-variants.vcf.gz" % os.path.splitext(align_bams[0])[0]
if not utils.file_exists(out_file):
with file_transaction(config, out_file) as tx_out_file:
if len(align_bams) > 1:
message = ("Scalpel does not currently support batch calling!")
raise ValueError(message)
input_bams = " ".join("%s" % x for x in align_bams)
tmp_path = "%s-scalpel-work" % utils.splitext_plus(out_file)[0]
tx_tmp_path = "%s-scalpel-work" % utils.splitext_plus(
tx_out_file)[0]
if os.path.exists(tmp_path):
utils.remove_safe(tmp_path)
opts = " ".join(
_scalpel_options_from_config(items, config, out_file, region,
tmp_path))
opts += " --dir %s" % tx_tmp_path
min_cov = "3" # minimum coverage
opts += " --mincov %s" % min_cov
perl_exports = utils.get_perl_exports(os.path.dirname(tx_out_file))
cmd = (
"{perl_exports} && "
"scalpel-discovery --single {opts} --ref {ref_file} --bam {input_bams} "
)
do.run(cmd.format(**locals()), "Genotyping with Scalpel", {})
shutil.move(tx_tmp_path, tmp_path)
# parse produced variant file further
scalpel_tmp_file = bgzip_and_index(
os.path.join(tmp_path, "variants.indel.vcf"), config)
compress_cmd = "| bgzip -c" if out_file.endswith("gz") else ""
bcftools_cmd_chi2 = get_scalpel_bcftools_filter_expression(
"chi2", config)
sample_name_str = items[0]["name"][1]
fix_ambig = vcfutils.fix_ambiguous_cl()
cl2 = (
"{bcftools_cmd_chi2} {scalpel_tmp_file} | "
r"sed 's/FORMAT\tsample\(_name\)\{{0,1\}}/FORMAT\t{sample_name_str}/g' "
"| {fix_ambig} | vcfallelicprimitives -t DECOMPOSED --keep-geno | vcffixup - | vcfstreamsort "
"{compress_cmd} > {tx_out_file}")
do.run(cl2.format(**locals()), "Finalising Scalpel variants", {})
ann_file = annotation.annotate_nongatk_vcf(out_file, align_bams,
assoc_files.get("dbsnp"),
ref_file, config)
return ann_file
def _run_workflow(items, paired, workflow_file, work_dir):
"""Run manta analysis inside prepared workflow directory.
"""
data = paired.tumor_data if paired else items[0]
out_file = os.path.join(work_dir, "results", "variants",
"somaticSV.vcf.gz" if paired and paired.normal_bam else "diploidSV.vcf.gz")
if not utils.file_exists(out_file):
utils.remove_safe(os.path.join(work_dir, "workspace"))
cmd = [sys.executable, workflow_file, "-m", "local", "-j", dd.get_num_cores(data),
"--quiet"]
do.run(cmd, "Run manta SV analysis")
utils.remove_safe(os.path.join(work_dir, "workspace"))
return out_file
def _grabix_index(data):
"""Create grabix index of bgzip input file.
grabix does not allow specification of output file, so symlink the original
file into a transactional directory.
"""
in_file = data["bgzip_file"]
config = data["config"]
grabix = config_utils.get_program("grabix", config)
gbi_file = in_file + ".gbi"
if tz.get_in(["algorithm", "align_split_size"], config) is not False:
if not utils.file_exists(gbi_file) or _is_partial_index(gbi_file):
utils.remove_safe(gbi_file)
with file_transaction(data, gbi_file) as tx_gbi_file:
tx_in_file = os.path.splitext(tx_gbi_file)[0]
utils.symlink_plus(in_file, tx_in_file)
do.run([grabix, "index", tx_in_file], "Index input with grabix: %s" % os.path.basename(in_file))
assert utils.file_exists(gbi_file)
return [gbi_file]
def tabix_index(in_file, config, preset=None, tabix_args=None):
"""Index a file using tabix.
"""
in_file = os.path.abspath(in_file)
out_file = in_file + ".tbi"
if not utils.file_exists(out_file) or not utils.file_uptodate(out_file, in_file):
# Remove old index files to prevent linking into tx directory
utils.remove_safe(out_file)
with file_transaction(config, out_file) as tx_out_file:
tabix = tools.get_tabix_cmd(config)
tx_in_file = os.path.splitext(tx_out_file)[0]
utils.symlink_plus(in_file, tx_in_file)
if tabix_args:
cmd = "{tabix} -f {tabix_args} {tx_in_file}"
else:
preset = _guess_preset(in_file) if preset is None else preset
cmd = "{tabix} -f -p {preset} {tx_in_file}"
do.run(cmd.format(**locals()), "tabix index %s" % os.path.basename(in_file))
return out_file
def _run_workflow(items, paired, workflow_file, work_dir):
"""Run manta analysis inside prepared workflow directory.
"""
data = paired.tumor_data if paired else items[0]
if paired:
if paired.normal_bam:
base_file = "somaticSV.vcf.gz"
else:
base_file = "tumorSV.vcf.gz"
else:
base_file = "diploidSV.vcf.gz"
out_file = os.path.join(work_dir, "results", "variants", base_file)
if not utils.file_exists(out_file):
utils.remove_safe(os.path.join(work_dir, "workspace"))
cmd = [sys.executable, workflow_file, "-m", "local", "-j", dd.get_num_cores(data),
]
do.run(cmd, "Run manta SV analysis")
utils.remove_safe(os.path.join(work_dir, "workspace"))
return out_file
def _grabix_index(data):
"""Create grabix index of bgzip input file.
grabix does not allow specification of output file, so symlink the original
file into a transactional directory.
"""
in_file = data["bgzip_file"]
config = data["config"]
grabix = config_utils.get_program("grabix", config)
gbi_file = in_file + ".gbi"
# We always build grabix input so we can use it for counting reads and doing downsampling
if not utils.file_exists(gbi_file) or _is_partial_index(gbi_file):
utils.remove_safe(gbi_file)
with file_transaction(data, gbi_file) as tx_gbi_file:
tx_in_file = os.path.splitext(tx_gbi_file)[0]
utils.symlink_plus(in_file, tx_in_file)
do.run([grabix, "index", tx_in_file], "Index input with grabix: %s" % os.path.basename(in_file))
assert utils.file_exists(gbi_file)
return [gbi_file]
def _run_scalpel_caller(align_bams, items, ref_file, assoc_files,
region=None, out_file=None):
"""Detect indels with Scalpel.
Single sample mode.
"""
config = items[0]["config"]
if out_file is None:
out_file = "%s-variants.vcf.gz" % os.path.splitext(align_bams[0])[0]
if not utils.file_exists(out_file):
with file_transaction(config, out_file) as tx_out_file:
if len(align_bams) > 1:
message = ("Scalpel does not currently support batch calling!")
raise ValueError(message)
input_bams = " ".join("%s" % x for x in align_bams)
tmp_path = "%s-scalpel-work" % utils.splitext_plus(out_file)[0]
tx_tmp_path = "%s-scalpel-work" % utils.splitext_plus(tx_out_file)[0]
if os.path.exists(tmp_path):
utils.remove_safe(tmp_path)
opts = " ".join(_scalpel_options_from_config(items, config, out_file, region, tmp_path))
opts += " --dir %s" % tx_tmp_path
min_cov = "3" # minimum coverage
opts += " --mincov %s" % min_cov
perl_exports = utils.get_perl_exports(os.path.dirname(tx_out_file))
cmd = ("{perl_exports} && "
"scalpel-discovery --single {opts} --ref {ref_file} --bam {input_bams} ")
do.run(cmd.format(**locals()), "Genotyping with Scalpel", {})
shutil.move(tx_tmp_path, tmp_path)
# parse produced variant file further
scalpel_tmp_file = bgzip_and_index(os.path.join(tmp_path, "variants.indel.vcf"), config)
compress_cmd = "| bgzip -c" if out_file.endswith("gz") else ""
bcftools_cmd_chi2 = get_scalpel_bcftools_filter_expression("chi2", config)
sample_name_str = items[0]["name"][1]
fix_ambig = vcfutils.fix_ambiguous_cl()
cl2 = ("{bcftools_cmd_chi2} {scalpel_tmp_file} | "
r"sed 's/FORMAT\tsample\(_name\)\{{0,1\}}/FORMAT\t{sample_name_str}/g' "
"| {fix_ambig} | vcfallelicprimitives -t DECOMPOSED --keep-geno | vcffixup - | vcfstreamsort "
"{compress_cmd} > {tx_out_file}")
do.run(cl2.format(**locals()), "Finalising Scalpel variants", {})
ann_file = annotation.annotate_nongatk_vcf(out_file, align_bams,
assoc_files.get("dbsnp"),
ref_file, config)
return ann_file
def tabix_index(in_file, config, preset=None, tabix_args=None):
"""Index a file using tabix.
"""
in_file = os.path.abspath(in_file)
out_file = in_file + ".tbi"
if not utils.file_exists(out_file) or not utils.file_uptodate(out_file, in_file):
# Remove old index files to prevent linking into tx directory
utils.remove_safe(out_file)
with file_transaction(config, out_file) as tx_out_file:
tabix = tools.get_tabix_cmd(config)
tx_in_file = os.path.splitext(tx_out_file)[0]
utils.symlink_plus(in_file, tx_in_file)
if tabix_args:
cmd = "{tabix} -f {tabix_args} {tx_in_file}"
else:
preset = _guess_preset(in_file) if preset is None else preset
cmd = "{tabix} -f -p {preset} {tx_in_file}"
do.run(cmd.format(**locals()), "tabix index %s" % os.path.basename(in_file))
return out_file
def _rename_allelic_fraction_field(orig_file,config):
"""Rename allelic fraction field in mutect output
from FA to FREQ to standarize with other tools
"""
tmp_file = orig_file.replace(".vcf.gz","-fix.vcf")
with file_transaction(tmp_file) as tx_in_file:
with open(tx_in_file,'w') as out_handle:
with open_gzipsafe(orig_file) as handle:
for line in handle:
if line.startswith("##FORMAT=<ID=FA"):
line = line.replace("=FA","=FREQ")
if not line.startswith("#"):
line = line.replace("FA","FREQ")
out_handle.write(line)
out_file = orig_file.replace(".gz","")
remove_safe(orig_file)
shutil.move(tmp_file, out_file )
with open(tmp_file, "w") as out_handle:
out_handle.write("Moved to {0}".format(out_file))
out_file = bgzip_and_index(out_file, config)
def _rename_allelic_fraction_field(orig_file, config):
"""Rename allelic fraction field in mutect output
from FA to FREQ to standarize with other tools
"""
tmp_file = orig_file.replace(".vcf.gz", "-fix.vcf")
with file_transaction(config, tmp_file) as tx_in_file:
with open(tx_in_file, 'w') as out_handle:
with open_gzipsafe(orig_file) as handle:
for line in handle:
if line.startswith("##FORMAT=<ID=FA"):
line = line.replace("=FA", "=FREQ")
if not line.startswith("#"):
line = line.replace("FA", "FREQ")
out_handle.write(line)
out_file = orig_file.replace(".gz", "")
remove_safe(orig_file)
shutil.move(tmp_file, out_file)
with open(tmp_file, "w") as out_handle:
out_handle.write("Moved to {0}".format(out_file))
out_file = bgzip_and_index(out_file, config)
def _install_kraken_db(datadir, args):
"""Install kraken minimal DB in genome folder.
"""
import requests
kraken = os.path.join(datadir, "genomes/kraken")
url = "https://ccb.jhu.edu/software/kraken/dl/minikraken.tgz"
compress = os.path.join(kraken, os.path.basename(url))
base, ext = utils.splitext_plus(os.path.basename(url))
db = os.path.join(kraken, base)
tooldir = args.tooldir or get_defaults()["tooldir"]
requests.packages.urllib3.disable_warnings()
last_mod = urllib.request.urlopen(url).info().get('Last-Modified')
last_mod = dateutil.parser.parse(last_mod).astimezone(dateutil.tz.tzutc())
if os.path.exists(os.path.join(tooldir, "bin", "kraken")):
if not os.path.exists(db):
is_new_version = True
else:
cur_file = glob.glob(os.path.join(kraken, "minikraken_*"))[0]
cur_version = datetime.datetime.utcfromtimestamp(
os.path.getmtime(cur_file))
is_new_version = last_mod.date() > cur_version.date()
if is_new_version:
shutil.move(cur_file, cur_file.replace('minikraken', 'old'))
if not os.path.exists(kraken):
utils.safe_makedir(kraken)
if is_new_version:
if not os.path.exists(compress):
subprocess.check_call(
["wget", "-O", compress, url, "--no-check-certificate"])
cmd = ["tar", "-xzvf", compress, "-C", kraken]
subprocess.check_call(cmd)
last_version = glob.glob(os.path.join(kraken, "minikraken_*"))
utils.symlink_plus(os.path.join(kraken, last_version[0]),
os.path.join(kraken, "minikraken"))
utils.remove_safe(compress)
else:
print("You have the latest version %s." % last_mod)
else:
raise argparse.ArgumentTypeError(
"kraken not installed in tooldir %s." %
os.path.join(tooldir, "bin", "kraken"))
def tx_tmpdir(data=None, base_dir=None, remove=True):
"""Context manager to create and remove a transactional temporary directory.
Handles creating a transactional directory for running commands in. Will
use either the current directory or /tmp/bcbiotx.
Creates an intermediary location and time specific directory for global
temporary directories to prevent collisions.
data can be the full world information object being process or a
configuration dictionary.
"""
base_dir = base_dir or os.getcwd()
tmpdir_base = utils.get_abspath(_get_base_tmpdir(data, base_dir))
utils.safe_makedir(tmpdir_base)
tmp_dir = tempfile.mkdtemp(dir=tmpdir_base)
logger.debug("Created tmp dir %s " % tmp_dir)
try:
yield tmp_dir
finally:
if remove:
utils.remove_safe(tmp_dir)
def _install_kraken_db(datadir, args):
"""Install kraken minimal DB in genome folder.
"""
kraken = os.path.join(datadir, "genomes/kraken")
url = "https://ccb.jhu.edu/software/kraken/dl/minikraken.tgz"
compress = os.path.join(kraken, os.path.basename(url))
base, ext = utils.splitext_plus(os.path.basename(url))
db = os.path.join(kraken, base)
tooldir = args.tooldir or get_defaults()["tooldir"]
if os.path.exists(os.path.join(tooldir, "bin", "kraken")):
if not os.path.exists(kraken):
utils.safe_makedir(kraken)
if not os.path.exists(db):
if not os.path.exists(compress):
subprocess.check_call(["wget", "-O", compress, url, "--no-check-certificate"])
cmd = ["tar", "-xzvf", compress, "-C", kraken]
subprocess.check_call(cmd)
shutil.move(os.path.join(kraken, "minikraken_20140330"), os.path.join(kraken, "minikraken"))
utils.remove_safe(compress)
else:
raise argparse.ArgumentTypeError("kraken not installed in tooldir %s." %
os.path.join(tooldir, "bin", "kraken"))
def _move_tmp_files(safe, orig):
exts = {
".vcf": ".idx",
".bam": ".bai",
".vcf.gz": ".tbi",
".bed.gz": ".tbi"
}
utils.safe_makedir(os.path.dirname(orig))
# If we are rolling back a directory and it already exists
# this will avoid making a nested set of directories
if os.path.isdir(orig) and os.path.isdir(safe):
utils.remove_safe(orig)
_move_file_with_sizecheck(safe, orig)
# Move additional, associated files in the same manner
for check_ext, check_idx in exts.iteritems():
if not safe.endswith(check_ext):
continue
safe_idx = safe + check_idx
if os.path.exists(safe_idx):
_move_file_with_sizecheck(safe_idx, orig + check_idx)
def tx_tmpdir(data=None, base_dir=None, remove=True):
"""Context manager to create and remove a transactional temporary directory.
Handles creating a transactional directory for running commands in. Will
use either the current directory or /tmp/bcbiotx.
Creates an intermediary location and time specific directory for global
temporary directories to prevent collisions.
data can be the full world information object being process or a
configuration dictionary.
"""
base_dir = base_dir or os.getcwd()
tmpdir_base = utils.get_abspath(_get_base_tmpdir(data, base_dir))
utils.safe_makedir(tmpdir_base)
tmp_dir = tempfile.mkdtemp(dir=tmpdir_base)
#logger.debug("Created tmp dir %s " % tmp_dir)
try:
yield tmp_dir
finally:
if remove:
utils.remove_safe(tmp_dir)
def _move_tmp_files(safe, orig):
exts = {
".vcf": ".idx",
".bam": ".bai",
".vcf.gz": ".tbi",
".bed.gz": ".tbi"
}
utils.safe_makedir(os.path.dirname(orig))
# If we are rolling back a directory and it already exists
# this will avoid making a nested set of directories
if os.path.isdir(orig) and os.path.isdir(safe):
utils.remove_safe(orig)
_move_file_with_sizecheck(safe, orig)
# Move additional, associated files in the same manner
for check_ext, check_idx in exts.items():
if not safe.endswith(check_ext):
continue
safe_idx = safe + check_idx
if os.path.exists(safe_idx):
_move_file_with_sizecheck(safe_idx, orig + check_idx)
def run(name, chip_bam, rep_bam, input_bam, gtf_file, out_dir, rlength, rpair, config):
"""
Run rmats for muatant and control samples avoiding
errors due to samples.
"""
# output file name need to have the caller name
MATS_output = os.path.join(out_dir, name + "_MATS_output")
MATS_dir = os.path.join(out_dir, "MATS_output")
rmats_file = os.path.join(out_dir, "summary.txt")
out_file = os.path.join(out_dir, name + "_summary.txt")
'''myCmd = 'samtools view '+chip_bam+' | head -n 1'
status,output=commands.getstatusoutput(myCmd)
rlength=len(output.strip().split('\t')[9])'''
libType = _get_stranded_flag(config)
if rep_bam != "":
chip_bam = chip_bam + "," + rep_bam
if utils.file_exists(out_file):
return out_file
rmats = config_utils.get_program("rmats", config)
options = " ".join(config_utils.get_resources("rmats", config).get("options", ""))
with utils.chdir(out_dir):
cmd = _rmats_cmd()
try:
do.run(cmd.format(**locals()), "rmats for %s" % name)
utils.move_safe(rmats_file, out_file)
utils.move_safe(MATS_dir, MATS_output)
repdir_dir = os.path.join(out_dir,"SAMPLE_1")
utils.remove_safe(repdir_dir)
repdir_dir = os.path.join(out_dir,"SAMPLE_2")
utils.remove_safe(repdir_dir)
print repdir_dir
except subprocess.CalledProcessError:
raise RuntimeWarning("rMATS terminated with an error.\n"
"Please, check the message and report "
"error if it is related to bcbio.\n"
"You can add specific options for the sample "
"setting resources as explained in docs: "
"https://bcbio-nextgen.readthedocs.org/en/latest/contents/configuration.html#sample-specific-resources")
return (out_file)
def index(in_bam, config, check_timestamp=True):
"""Index a BAM file, skipping if index present.
Centralizes BAM indexing providing ability to switch indexing approaches.
"""
assert is_bam(in_bam), "%s in not a BAM file" % in_bam
index_file = "%s.bai" % in_bam
alt_index_file = "%s.bai" % os.path.splitext(in_bam)[0]
if check_timestamp:
bai_exists = utils.file_uptodate(index_file, in_bam) or utils.file_uptodate(alt_index_file, in_bam)
else:
bai_exists = utils.file_exists(index_file) or utils.file_exists(alt_index_file)
if not bai_exists:
# Remove old index files and re-run to prevent linking into tx directory
for fname in [index_file, alt_index_file]:
utils.remove_safe(fname)
samtools = config_utils.get_program("samtools", config)
num_cores = config["algorithm"].get("num_cores", 1)
with file_transaction(config, index_file) as tx_index_file:
cmd = "{samtools} index -@ {num_cores} {in_bam} {tx_index_file}"
do.run(cmd.format(**locals()), "Index BAM file: %s" % os.path.basename(in_bam))
return index_file if utils.file_exists(index_file) else alt_index_file
def index(in_bam, config, check_timestamp=True):
"""Index a BAM file, skipping if index present.
Centralizes BAM indexing providing ability to switch indexing approaches.
"""
assert is_bam(in_bam), "%s in not a BAM file" % in_bam
index_file = "%s.bai" % in_bam
alt_index_file = "%s.bai" % os.path.splitext(in_bam)[0]
if check_timestamp:
bai_exists = utils.file_uptodate(
index_file, in_bam) or utils.file_uptodate(alt_index_file, in_bam)
else:
bai_exists = utils.file_exists(index_file) or utils.file_exists(
alt_index_file)
if not bai_exists:
# Remove old index files and re-run to prevent linking into tx directory
for fname in [index_file, alt_index_file]:
utils.remove_safe(fname)
sambamba = _get_sambamba(config)
assert sambamba, "Did not find sambamba for indexing"
samtools = config_utils.get_program("samtools", config)
num_cores = config["algorithm"].get("num_cores", 1)
with file_transaction(config, index_file) as tx_index_file:
assert tx_index_file.find(".bam.bai") > 0
tx_bam_file = tx_index_file.replace(".bam.bai", ".bam")
utils.symlink_plus(in_bam, tx_bam_file)
try:
cmd = "{sambamba} index -t {num_cores} {tx_bam_file}"
do.run(
cmd.format(**locals()),
"Index BAM file with sambamba: %s" %
os.path.basename(in_bam))
except subprocess.CalledProcessError:
cmd = "{samtools} index {in_bam} {tx_index_file}"
do.run(
cmd.format(**locals()),
"Backup single thread index of BAM file with samtools: %s"
% os.path.basename(in_bam))
return index_file if utils.file_exists(index_file) else alt_index_file
