def _run_fastqc(global_config, sample_config, sorted_libraries_by_insert):
mainDir = os.getcwd()
FastqcFolder = os.path.join(os.getcwd(), "fastqc")
if not os.path.exists(FastqcFolder):
os.makedirs(FastqcFolder)
program=global_config["Tools"]["fastqc"]["bin"]
program_options=global_config["Tools"]["fastqc"]["options"]
for library, libraryInfo in sorted_libraries_by_insert:
command = [program]
for option in program_options:
command.append(option)
read1=libraryInfo["pair1"]
read2=libraryInfo["pair2"]
command.append(read1)
if read2 is not None:
command.append(read2)
common.print_command(command)
sample_config["commands"] += "\n" + common.get_command_str(command)
folder_output_name = os.path.join(FastqcFolder,
os.path.basename(read1).split(".fastq.gz")[0])
if not common.check_dryrun(sample_config) and not \
os.path.exists("{}_fastqc.zip".format(folder_output_name)):
fastq_stdOut = open(os.path.join(FastqcFolder,
"{}_fastqc.stdout".format(library)), "a")
fastq_stdErr = open(os.path.join(FastqcFolder,
"{}_fastqc.stderr".format(library)), "a")
subprocess.call(command, stdout=fastq_stdOut, stderr=fastq_stdErr)
sample_config["fastqc"] = FastqcFolder
return sample_config
def _run_fastqc(global_config, sample_config, sorted_libraries_by_insert):
mainDir = os.getcwd()
FastqcFolder = os.path.join(os.getcwd(), "fastqc")
if not os.path.exists(FastqcFolder):
os.makedirs(FastqcFolder)
program = global_config["Tools"]["fastqc"]["bin"]
program_options = global_config["Tools"]["fastqc"]["options"]
for library, libraryInfo in sorted_libraries_by_insert:
command = [program]
for option in program_options:
command.append(option)
read1 = libraryInfo["pair1"]
read2 = libraryInfo["pair2"]
command.append(read1)
if read2 is not None:
command.append(read2)
common.print_command(command)
sample_config["commands"] += "\n" + common.get_command_str(command)
folder_output_name = os.path.join(
FastqcFolder,
os.path.basename(read1).split(".fastq.gz")[0])
if not common.check_dryrun(sample_config) and not \
os.path.exists("{}_fastqc.zip".format(folder_output_name)):
fastq_stdOut = open(
os.path.join(FastqcFolder, "{}_fastqc.stdout".format(library)),
"a")
fastq_stdErr = open(
os.path.join(FastqcFolder, "{}_fastqc.stderr".format(library)),
"a")
subprocess.call(command, stdout=fastq_stdOut, stderr=fastq_stdErr)
sample_config["fastqc"] = FastqcFolder
return sample_config
def _run_abyss(global_config, sample_config, sorted_libraries_by_insert):
mainDir = os.getcwd()
ABySS_Kmer_Folder = os.path.join(os.getcwd(), "abyss_kmer")
if "kmer" not in sample_config:
sys.exit("error in _run_abyss QCcontrol: kmer must be present in \
sample_config.yaml")
kmer = sample_config["kmer"]
if not os.path.exists(ABySS_Kmer_Folder):
os.makedirs(ABySS_Kmer_Folder)
os.chdir(ABySS_Kmer_Folder)
program = global_config["Tools"]["abyss"]["bin"]
program = os.path.join(os.path.dirname(program), "ABYSS-P")
program_options=global_config["Tools"]["abyss"]["options"]
if "abyss" in sample_config:
program_options=sample_config["abyss"]
threads = 16 # default for UPPMAX
if "threads" in sample_config :
threads = sample_config["threads"]
command = "mpirun -np {} {} ".format(threads, program)
command += "-k {} ".format(kmer)
command += "--coverage-hist=histogram.hist -o preUnitgs.fa"
for library, libraryInfo in sorted_libraries_by_insert:
read1=libraryInfo["pair1"]
read2=libraryInfo["pair2"]
orientation = libraryInfo["orientation"]
if orientation=="innie" or orientation=="outtie":
command += " {} ".format(read1)
if read2 is not None:
command += " {} ".format(read2)
if orientation == "none":
command += " {} ".format(read1)
common.print_command(command)
sample_config["commands"] += "\n" + common.get_command_str(command)
if not common.check_dryrun(sample_config) and not \
os.path.exists("histogram.hist"):
ABySS_Kmer_stdOut = open("ABySS_Kmer_Folder.stdOut", "a")
ABySS_Kmer_stdErr = open("ABySS_Kmer_Folder.stdErr", "a")
returnValue = subprocess.call(command, shell=True, \
stdout=ABySS_Kmer_stdOut, stderr=ABySS_Kmer_stdErr)
if returnValue > 0:
print("ABySS kmer plotting failed: unkwnown reason")
else :
subprocess.call(("rm", "preUnitgs.fa"))
_plotKmerFixed(1,200, kmer, "kmer_coverage_1_200.png")
_plotKmerFixed(1,500, kmer, "kmer_coverage_1_500.png")
_plotKmerFixed(15,200, kmer, "kmer_coverage_15_200.png")
_plotKmerFixed(15,500, kmer, "kmer_coverage_15_500.png")
_plotKmer(kmer, "kmer_coverage.png")
os.chdir("..")
sample_config["abyss"] = ABySS_Kmer_Folder
return sample_config
def _run_abyss(global_config, sample_config, sorted_libraries_by_insert):
mainDir = os.getcwd()
ABySS_Kmer_Folder = os.path.join(os.getcwd(), "abyss_kmer")
if "kmer" not in sample_config:
sys.exit("error in _run_abyss QCcontrol: kmer must be present in \
sample_config.yaml")
kmer = sample_config["kmer"]
if not os.path.exists(ABySS_Kmer_Folder):
os.makedirs(ABySS_Kmer_Folder)
os.chdir(ABySS_Kmer_Folder)
program = global_config["Tools"]["abyss"]["bin"]
program = os.path.join(os.path.dirname(program), "ABYSS-P")
program_options = global_config["Tools"]["abyss"]["options"]
if "abyss" in sample_config:
program_options = sample_config["abyss"]
threads = 16 # default for UPPMAX
if "threads" in sample_config:
threads = sample_config["threads"]
command = "mpirun -np {} {} ".format(threads, program)
command += "-k {} ".format(kmer)
command += "--coverage-hist=histogram.hist -o preUnitgs.fa"
for library, libraryInfo in sorted_libraries_by_insert:
read1 = libraryInfo["pair1"]
read2 = libraryInfo["pair2"]
orientation = libraryInfo["orientation"]
if orientation == "innie" or orientation == "outtie":
command += " {} ".format(read1)
if read2 is not None:
command += " {} ".format(read2)
if orientation == "none":
command += " {} ".format(read1)
common.print_command(command)
sample_config["commands"] += "\n" + common.get_command_str(command)
if not common.check_dryrun(sample_config) and not \
os.path.exists("histogram.hist"):
ABySS_Kmer_stdOut = open("ABySS_Kmer_Folder.stdOut", "a")
ABySS_Kmer_stdErr = open("ABySS_Kmer_Folder.stdErr", "a")
returnValue = subprocess.call(command, shell=True, \
stdout=ABySS_Kmer_stdOut, stderr=ABySS_Kmer_stdErr)
if returnValue > 0:
print("ABySS kmer plotting failed: unkwnown reason")
else:
subprocess.call(("rm", "preUnitgs.fa"))
_plotKmerFixed(1, 200, kmer, "kmer_coverage_1_200.png")
_plotKmerFixed(1, 500, kmer, "kmer_coverage_1_500.png")
_plotKmerFixed(15, 200, kmer, "kmer_coverage_15_200.png")
_plotKmerFixed(15, 500, kmer, "kmer_coverage_15_500.png")
_plotKmer(kmer, "kmer_coverage.png")
os.chdir("..")
sample_config["abyss"] = ABySS_Kmer_Folder
return sample_config
def _run_kmergenie(global_config, sample_config, sorted_libraries_by_insert):
"""Runs kmergenie to establish a recommended kmer size for assembly"""
maindir = os.getcwd()
kmerdir = os.path.join(maindir, "kmergenie")
if not os.path.exists(kmerdir):
os.makedirs(kmerdir)
os.chdir(kmerdir)
#Write a list of input fastq files for kmergenie
kmer_input = os.path.join(kmerdir,
"{}kmerinput.txt".format(sample_config.get("output","")))
program = global_config["Tools"]["kmergenie"]["bin"]
program_options=global_config["Tools"]["kmergenie"]["options"]
# Could be useful to add --diploid if sample is highly heterozygous
if "kmergenie" in sample_config:
program_options=sample_config["kmergenie"]
threads = "" # Kmergenie will spawn number_of_cores - 1 threads by default
if "threads" in sample_config :
threads = sample_config["threads"]
cmd_list = [program, kmer_input]
for option in filter(None, program_options):
cmd_list.append(option)
if threads:
cmd_list.append("-t {}".format(threads))
command = " ".join(cmd_list)
common.print_command(command)
sample_config["commands"] += "\n" + common.get_command_str(command)
if not common.check_dryrun(sample_config):
with open(kmer_input, "w") as f:
for lib, lib_info in sorted_libraries_by_insert:
f.write(lib_info["pair1"] + "\n")
f.write(lib_info["pair2"] + "\n")
stdOut = open("kmergenie.stdOut", "w")
stdErr = open("kmergenie.stdErr", "w")
returnValue = subprocess.call(cmd_list, stdout=stdOut, stderr=stdErr)
if returnValue != 0:
print("error while running command: {}".format(command))
else:
_kmergenie_plot("histograms.dat")
sample_config["kmergenie"] = kmerdir
os.chdir(maindir)
return sample_config
def _run_kmergenie(global_config, sample_config, sorted_libraries_by_insert):
"""Runs kmergenie to establish a recommended kmer size for assembly"""
maindir = os.getcwd()
kmerdir = os.path.join(maindir, "kmergenie")
if not os.path.exists(kmerdir):
os.makedirs(kmerdir)
os.chdir(kmerdir)
#Write a list of input fastq files for kmergenie
kmer_input = os.path.join(kmerdir,
"{}kmerinput.txt".format(sample_config.get("output","")))
program = global_config["Tools"]["kmergenie"]["bin"]
program_options=global_config["Tools"]["kmergenie"]["options"]
# Could be useful to add --diploid if sample is highly heterozygous
if "kmergenie" in sample_config:
program_options=sample_config["kmergenie"]
threads = "" # Kmergenie will spawn number_of_cores - 1 threads by default
if "threads" in sample_config :
threads = sample_config["threads"]
cmd_list = [program, kmer_input]
for option in filter(None, program_options):
cmd_list.append(option)
if threads:
cmd_list.append("-t {}".format(threads))
command = " ".join(cmd_list)
common.print_command(command)
sample_config["commands"] += "\n" + common.get_command_str(command)
if not common.check_dryrun(sample_config):
with open(kmer_input, "w") as f:
for lib, lib_info in sorted_libraries_by_insert:
f.write(lib_info["pair1"] + "\n")
f.write(lib_info["pair2"] + "\n")
stdOut = open("kmergenie.stdOut", "w")
stdErr = open("kmergenie.stdErr", "w")
returnValue = subprocess.call(cmd_list, stdout=stdOut, stderr=stdErr)
if returnValue != 0:
print("error while running command: {}".format(command))
else:
_kmergenie_plot("histograms.dat")
sample_config["kmergenie"] = kmerdir
os.chdir(maindir)
return sample_config
def _run_trimmomatic(global_config, sample_config, sorted_libraries_by_insert):
program = global_config["Tools"]["trimmomatic"]["bin"]
program_folder = os.path.dirname(program)
if "adapters" not in sample_config:
sys.exit("running MP pipeline, adapters file to be used in trimming"
"are needed for Trimmomatic. Please specify them"
"in the sample configuration file and rerun")
adapterFile = sample_config["adapters"]
if not os.path.exists(adapterFile):
sys.exit("Trimmomatic cannot be run as adapter file is not specified"
"or points to unknown position: {}".format(adapterFile))
mainDirectory = os.getcwd()
trimmomaticDir = os.path.join(mainDirectory, "Trimmomatic")
if not os.path.exists(trimmomaticDir):
os.makedirs(trimmomaticDir)
os.chdir(trimmomaticDir)
#now I am in running dir, I need to process one by one the libraries
threads = 8
if "threads" in sample_config:
threads = sample_config["threads"]
for library, libraryInfo in sorted_libraries_by_insert:
read1=libraryInfo["pair1"]
read2=libraryInfo["pair2"]
orientation = libraryInfo["orientation"]
if read2 is not None:
read1_baseName = os.path.split(read1)[1].split(".")[0]
read2_baseName = os.path.split(read2)[1].split(".")[0]
output_read1_pair = os.path.join(trimmomaticDir,
"{}.fastq.gz".format(read1_baseName))
output_read1_sing = os.path.join(trimmomaticDir,
"{}_u.fastq.gz".format(read1_baseName))
output_read2_pair = os.path.join(trimmomaticDir,
"{}.fastq.gz".format(read2_baseName))
output_read2_sing = os.path.join(trimmomaticDir,
"{}_u.fastq.gz".format(read2_baseName))
command = ["java", "-jar", program, "PE", "-threads",
"{}".format(threads), "-phred33", read1, read2,
output_read1_pair, output_read1_sing, output_read2_pair,
output_read2_sing,
"ILLUMINACLIP:{}:2:30:10".format(adapterFile),
"LEADING:3", "TRAILING:3", "SLIDINGWINDOW:4:15",
"MINLEN:30"]
common.print_command(command)
sample_config["commands"] += "\n" + common.get_command_str(command)
# do not execute is files have been already gennerated
if not common.check_dryrun(sample_config) and not \
os.path.exists(output_read1_pair):
stdOut = open("{}_trimmomatic.stdOut".format(read1_baseName),
"w")
stdErr = open("{}_trimmomatic.stdErr".format(read1_baseName),
"w")
returnValue = subprocess.call(command, stdout=stdOut,
stderr=stdErr) # run the program
if returnValue != 0:
print("error while running command: {}".format(command))
libraryInfo["pair1"] = output_read1_pair
libraryInfo["pair2"] = output_read2_pair
libraryInfo["trimmomatic"] = os.path.join(trimmomaticDir,
"{}_trimmomatic.stdErr".format(read1_baseName))
os.chdir(mainDirectory)
return sample_config
def _run_trimmomatic(global_config, sample_config, sorted_libraries_by_insert):
program = global_config["Tools"]["trimmomatic"]["bin"]
program_folder = os.path.dirname(program)
if "adapters" not in sample_config:
sys.exit("running MP pipeline, adapters file to be used in trimming"
"are needed for Trimmomatic. Please specify them"
"in the sample configuration file and rerun")
adapterFile = sample_config["adapters"]
if not os.path.exists(adapterFile):
sys.exit("Trimmomatic cannot be run as adapter file is not specified"
"or points to unknown position: {}".format(adapterFile))
mainDirectory = os.getcwd()
trimmomaticDir = os.path.join(mainDirectory, "Trimmomatic")
if not os.path.exists(trimmomaticDir):
os.makedirs(trimmomaticDir)
os.chdir(trimmomaticDir)
#now I am in running dir, I need to process one by one the libraries
threads = 8
if "threads" in sample_config:
threads = sample_config["threads"]
for library, libraryInfo in sorted_libraries_by_insert:
read1 = libraryInfo["pair1"]
read2 = libraryInfo["pair2"]
orientation = libraryInfo["orientation"]
if read2 is not None:
read1_baseName = os.path.split(read1)[1].split(".")[0]
read2_baseName = os.path.split(read2)[1].split(".")[0]
output_read1_pair = os.path.join(
trimmomaticDir, "{}.fastq.gz".format(read1_baseName))
output_read1_sing = os.path.join(
trimmomaticDir, "{}_u.fastq.gz".format(read1_baseName))
output_read2_pair = os.path.join(
trimmomaticDir, "{}.fastq.gz".format(read2_baseName))
output_read2_sing = os.path.join(
trimmomaticDir, "{}_u.fastq.gz".format(read2_baseName))
command = [
"java", "-jar", program, "PE", "-threads",
"{}".format(threads), "-phred33", read1, read2,
output_read1_pair, output_read1_sing, output_read2_pair,
output_read2_sing,
"ILLUMINACLIP:{}:2:30:10".format(adapterFile), "LEADING:3",
"TRAILING:3", "SLIDINGWINDOW:4:15", "MINLEN:30"
]
common.print_command(command)
sample_config["commands"] += "\n" + common.get_command_str(command)
# do not execute is files have been already gennerated
if not common.check_dryrun(sample_config) and not \
os.path.exists(output_read1_pair):
stdOut = open("{}_trimmomatic.stdOut".format(read1_baseName),
"w")
stdErr = open("{}_trimmomatic.stdErr".format(read1_baseName),
"w")
returnValue = subprocess.call(command,
stdout=stdOut,
stderr=stdErr) # run the program
if returnValue != 0:
print("error while running command: {}".format(command))
libraryInfo["pair1"] = output_read1_pair
libraryInfo["pair2"] = output_read2_pair
libraryInfo["trimmomatic"] = os.path.join(
trimmomaticDir, "{}_trimmomatic.stdErr".format(read1_baseName))
os.chdir(mainDirectory)
return sample_config
