def principal_component_analysis(exp):
out_dir = make_folder(f'{exp.scratch}PCA/')
bigwigs = {
sample: exp.sample_files[sample]['bw']
for sample in exp.samples if len(exp.sample_files[sample]['bw']) != 0
}
multibw_command = f"multiBigwigSummary bins -b {' '.join(list(bigwigs.values()))} -l {' '.join(list(bigwigs.keys()))} -p 4 --chromosomesToSkip chrM,chrX,chrY -o {out_dir}{exp.name}_bwsummary.npz"
correlation_command = f'plotCorrelation --corData {out_dir}{exp.name}_bwsummary.npz --corMethod pearson --whatToPlot heatmap --skipZeros --plotTitle "{exp.name} Binned Pearson Correlation Heatmap" --plotFileFormat png --outFileCorMatrix {out_dir}{exp.name}_CorMatrix.tab --colorMap Purples -o {out_dir}{exp.name}_CorHeatmap.png'
pca_command = f'plotPCA --corData {out_dir}{exp.name}_bwsummary.npz --plotTitle "{exp.name} PCA Plot" --plotFileFormat png --outFileNameData {out_dir}{exp.name}_PCA_data.tab --log2 -o {out_dir}{exp.name}_PCA_Plot.png'
command_list = [
submission_prepend(), multibw_command, correlation_command, pca_command
]
exp.job_id.append(
send_job(command_list=command_list,
job_name=f"{exp.name}_Cor_PCA",
job_log_folder=exp.job_folder,
q='general',
mem=4000,
log_file=exp.log_file,
project=exp.project,
cores=5,
run_main=exp.run_main))
exp.tasks_complete.append('PCA')
return exp
def preseq(exp):
output(
'\nRunning QC plots: library complexity extrapolation, signal correlation and pca plots.',
log_file=exp.log_file,
run_main=exp.run_main)
for sample in exp.samples:
out_dir = make_folder(f'{exp.scratch}QC/preseq/{sample}/')
command_list = [
submission_prepend(
f'preseq lc_extrap -bam -output {out_dir}{sample}_preseq.txt {exp.sample_files[sample]["bam"]}'
)
]
exp.job_id.append(
send_job(command_list=command_list,
job_name=f"{sample}_preseq",
job_log_folder=exp.job_folder,
q='general',
mem=5000,
log_file=exp.log_file,
project=exp.project,
cores=1,
run_main=exp.run_main))
exp.tasks_complete.append('preseq')
return exp
def fastq_screen(exp):
'''
Checks fastq files for contamination with alternative genomes using Bowtie2
'''
output(
f'Screening for contamination during sequencing: {datetime.now():%Y-%m-%d %H:%M:%S}\n',
log_file=exp.log_file,
run_main=exp.run_main)
# Make QC folder
exp.qc_folder = make_folder(f'{exp.scratch}QC/')
cwd = val_folder(os.getcwd())
os.chdir(exp.data_folder)
samples = [
file for file in exp.sample_df.Scratch_File1.tolist() if is_fastq(file)
]
# Submit fastqc and fastq_screen jobs for each sample
for sample in samples:
command_list = [
submission_prepend(
f'fastq_screen --threads 4 --aligner bowtie2 {sample}')
]
exp.job_id.append(
send_job(command_list=command_list,
job_name=f'{sample.split("/")[-1]}_fastq_screen',
job_log_folder=exp.job_folder,
q='general',
mem=3000,
log_file=exp.log_file,
project=exp.project,
cores=2,
run_main=exp.run_main))
time.sleep(1)
# Wait for jobs to finish
job_wait(exp.job_id, exp.log_file)
# move to qc folder
fastqs_files = glob.glob(f'{exp.data_folder}*screen*')
for f in fastqs_files:
copy2(f, exp.qc_folder)
os.remove(f)
# change to experimental directory in scratch
os.chdir(cwd)
exp.tasks_complete.append('Fastq_screen')
output(f'Screening complete: {datetime.now():%Y-%m-%d %H:%M:%S}\n',
log_file=exp.log_file,
run_main=exp.run_main)
return exp
def fastqc(exp):
'''
Performs fastq spec analysis with FastQC
'''
output('Assessing fastq quality. \n',
log_file=exp.log_file,
run_main=exp.run_main)
# Make QC folder
exp.qc_folder = make_folder(f'{exp.scratch}QC/')
all_samples = exp.sample_df.Scratch_File1.tolist(
) + exp.sample_df.Scratch_File2.tolist()
samples = [file for file in all_samples if is_fastq(file)]
for sample in samples:
command_list = [submission_prepend(f'fastqc {sample}')]
exp.job_id.append(
send_job(command_list=command_list,
job_name=f'{sample.split("/")[-1]}_fastqc',
job_log_folder=exp.job_folder,
q='general',
mem=5000,
log_file=exp.log_file,
project=exp.project,
run_main=exp.run_main))
# Wait for jobs to finish
job_wait(exp.job_id, exp.log_file)
# move to qc folder
fastqc_files = glob.glob(f'{exp.data_folder}*.zip')
fastqc_files = fastqc_files + glob.glob(f'{exp.data_folder}*.html')
for f in fastqc_files:
copy2(f, exp.qc_folder)
os.remove(f)
exp.tasks_complete.append('FastQC')
output(f'FastQC complete: {datetime.now():%Y-%m-%d %H:%M:%S}\n',
log_file=exp.log_file,
run_main=exp.run_main)
return exp
def trim(exp):
'''
Trimming based on standard UM SCCC Core Nextseq 500 technical errors.
Cudadapt can hard clip both ends, but may ignore 3' in future.
'''
output(f'Beginning fastq trimming: {datetime.now():%Y-%m-%d %H:%M:%S}\n',
log_file=exp.log_file,
run_main=exp.run_main)
for sample_dict in exp.sample_df[[
'Scratch_File1', 'Scratch_File2', 'Sequencer', 'Sample_Name'
]].to_dict(orient='records'):
quality = '--nextseq-trim=20' if sample_dict['Sequencer'].lower(
) == 'nextseq' else '-q 20'
seq_type = 'single' if sample_dict[
'Scratch_File2'] == 'none' else 'paired'
sample = sample_dict["Sample_Name"]
paired = f'{exp.data_folder}{sample}_trim_R2.fastq.gz'
single = f'{exp.data_folder}{sample}_trim.fastq.gz'
data_files = glob.glob(f'{exp.data_folder}*.gz')
if (single in data_files) or (paired in data_files):
continue
else:
output(f'Trimming {sample}: ',
log_file=exp.log_file,
run_main=exp.run_main)
if seq_type == 'paired':
cutadapt = f'cutadapt -j 4 -a AGATCGGAAGAGC -A AGATCGGAAGAGC --cores=10 {quality} -m 18 '
cutadapt += f'-o {exp.data_folder}{sample}_trim_R1.fastq.gz -p {exp.data_folder}{sample}_trim_R2.fastq.gz '
cutadapt += f'{sample_dict["Scratch_File1"]} {sample_dict["Scratch_File2"]}'
elif seq_type == 'single':
cutadapt = f'cutadapt -j 4 -a AGATCGGAAGAGC --cores=10 {quality} -m 18 '
cutadapt += f'-o {exp.data_folder}{sample}_trim_R1.fastq.gz {sample_dict["Scratch_File1"]}'
command_list = [submission_prepend(cutadapt)]
exp.job_id.append(
send_job(command_list=command_list,
job_name=f"{sample}_trim",
job_log_folder=exp.job_folder,
q='general',
mem=5000,
log_file=exp.log_file,
project=exp.project,
cores=2,
run_main=exp.run_main))
# Wait for jobs to finish
job_wait(exp.job_id, exp.log_file, exp.run_main)
# move logs to qc folder
output(
'\nTrimming logs are found in stdout files from bsub. Cutadapt does not handle log files in multi-core mode.',
log_file=exp.log_file,
run_main=exp.run_main)
exp.tasks_complete.append('Trim')
output(f'Trimming complete: {datetime.now():%Y-%m-%d %H:%M:%S}\n',
log_file=exp.log_file,
run_main=exp.run_main)
return exp
def spike(exp):
'''
If calling from jupyter. Change backend as needed.
Align sequencing files to drosophila.
'''
import pandas as pd
if len(exp.spike_samples) == 0:
output('Not processing Spike-ins',
log_file=exp.log_file,
run_main=exp.run_main)
exp.tasks_complete.append('Spike')
return exp
# Make QC folder
spike_folder = make_folder(f'{exp.scratch}spike/')
output('Processing samples with drosophila-spike in chromatin.',
log_file=exp.log_file,
run_main=exp.run_main)
for sample in exp.spike_samples:
bam = exp.sample_files[sample]['bam']
spike_command = [
submission_prepend(),
f'samtools view -b -f 4 {bam} | samtools sort -n - | samtools fastq - > {spike_folder}{sample}.bwa_unaligned.fastq',
f'bowtie2 -p 8 -x {exp.genome_indicies["spike_index"]} -U {spike_folder}{sample}.bwa_unaligned.fastq -S {spike_folder}{sample}.BDGP6.sam --very-sensitive-local -k 1 --no-unal',
f'samtools view -b -F 4 {spike_folder}{sample}.BDGP6.sam | samtools sort - > {spike_folder}{sample}.BDGP6.bam',
f'picard MarkDuplicates I={spike_folder}{sample}.BDGP6.bam O={spike_folder}{sample}.BDGP6.nodup.bam M={spike_folder}{sample}.BDGP6.nodups.markdups.qc ASSUME_SORTED=TRUE VALIDATION_STRINGENCY=LENIENT REMOVE_DUPLICATES=true',
f'samtools flagstat {spike_folder}{sample}.BDGP6.nodup.bam > {spike_folder}{sample}.unique_drosophila.flagstat.qc',
f'rm {spike_folder}{sample}.BDGP6.sam {spike_folder}{sample}.BDGP6.nodup.bam {spike_folder}{sample}*.fastq'
]
exp.job_id.append(
send_job(command_list=spike_command,
job_name=f"{sample}_spike",
job_log_folder=exp.job_folder,
q='general',
mem=10000,
log_file=exp.log_file,
project=exp.project,
cores=2,
run_main=exp.run_main))
# Wait for jobs to finish
job_wait(exp.job_id, exp.log_file, exp.run_main)
spike_reads = pd.DataFrame(index=['spike_reads', 'genome_reads'])
for sample in exp.spike_samples:
qc_file = f'{spike_folder}{sample}.unique_drosophila.flagstat.qc'
exp.sample_files[sample]['drosophila'] = qc_file
with open(qc_file, 'r') as fp:
spike_number = fp.read().split(' ')[0]
with open(exp.sample_files[sample]['nodup_flagstat']) as fp:
target_number = fp.read().split(' ')[0]
spike_reads[sample] = [spike_number, target_number]
exp.spike_reads = spike_reads.T
condition_dict = pd.Series(exp.sample_df.Condition.values,
index=exp.sample_df.Sample_Name).to_dict()
exp.spike_reads['Replicate'] = [
x.split('_')[-1] for x in exp.spike_reads.index.tolist()
]
exp.spike_reads['Condition'] = [
condition_dict[x] for x in exp.splike_reads.index.tolist()
]
for name, spike_conditions in exp.spike_comparisons.items():
out_dir = make_folder(f'{exp.scratch}spike/{name}')
plot = spike_in_plot(exp.spike_reads, spike_conditions, name, out_dir)
out_result(plot,
f'{name.replace("_", " ")} Spike-In Comparison',
run_main=exp.run_main)
output(
f'Spike-in comparison {name.replace("_", " ")} can be found here: {plot.replace(os.scratch, "")}'
)
output(f'Spike-in counts:\n {spike_reads.T}',
log_file=exp.log_file,
run_main=exp.run_main)
output('Spike-in alignment jobs finished.',
log_file=exp.log_file,
run_main=exp.run_main)
# Generate one dataframe for all spike_counts
output(
f"Spike-in processing complete: {datetime.now():%Y-%m-%d %H:%M:%S}\n",
log_file=exp.log_file,
run_main=exp.run_main)
exp.tasks_complete.append('Spike')
return exp
def UMI(exp):
# exp.data_type = 'bam'
IPs = exp.IPs
for experiment in IPs.Condition.unique().tolist():
UMI = True if 'yes' in IPs[IPs.Condition ==
experiment]['UMI'].tolist() else False
if not UMI:
return exp
else:
out_dir = make_folder(f'{exp.scratch}UMI/')
output('Deduplicating bam files using UMIs with UMI-tools.',
log_file=exp.log_file,
run_main=exp.run_main)
for index in IPs[IPs.Condition == experiment].index.tolist():
sample = IPs.loc[index, 'Sample_Name']
input_sample = IPs.loc[index, 'Background_Name']
bam = exp.sample_files[sample]['bam']
input_bam = exp.sample_files[input_sample]['bam']
nodup_bam = f'{out_dir}{sample}.UMI.dedup.bam'
nodup_input = f'{out_dir}{input_sample}.UMI.dedup.bam'
umi_string = 'umi_tools dedup --umi-separator=":" --output-stats={out_dir}{sample}deduplicated.qc -I {inbam} -S {outbam} -L {out_dir}{sample}.UMI.log'
seq_type = False if 'none' in IPs[
IPs.Condition ==
experiment]['Scratch_File2'].tolist() else True
if seq_type == 'paired':
umi_string += ' --paired'
command_list = [
submission_prepend(), f'samtools index {bam}',
f'samtools index {input_bam}',
umi_string.format(inbam=bam,
outbam=nodup_bam,
sample=sample,
out_dir=out_dir),
umi_string.format(inbam=input_bam,
outbam=nodup_input,
sample=input_sample,
out_dir=out_dir)
]
exp.job_id.append(
send_job(command_list=command_list,
job_name=f"{sample}_UMI_dedup",
job_log_folder=exp.job_folder,
q='bigmem',
mem=40000,
log_file=exp.log_file,
project=exp.project,
cores=1,
run_main=exp.run_main))
exp.sample_files[sample]['nodup_bam'] = nodup_bam
exp.sample_files[input_sample]['nodup_bam'] = nodup_input
job_wait(exp.job_id, exp.log_file)
output(
'Dedplication complete. Submitting deduplicated files for the remainder of processing.',
log_file=exp.log_file,
run_main=exp.run_main)
exp.tasks_complete.append('UMI')
return encode3(exp)
def encode3(exp):
if 'Stage' not in exp.tasks_complete:
output('Files not staged.\n', log_file=exp.log_file)
exp = stage(exp)
output('Running alignment and peak calling using ENCODE3 standards.',
log_file=exp.log_file,
run_main=exp.run_main)
output('ENCODE3 cromwell pipeline.',
log_file=exp.log_file,
run_main=exp.run_main)
out_dir = make_folder(f'{exp.scratch}ENCODE3/')
IPs = exp.IPs
end_types = {'q.gz': 'fastq', '.bam': 'bam'}
for experiment in IPs.Condition.unique().tolist():
exp_dir = make_folder(f'{out_dir}{experiment}/')
IP_sample_indicies = [(rep, index) for rep, index in enumerate(
IPs[IPs.Condition == experiment].index.tolist(), start=1)]
if len(IP_sample_indicies) > 6:
raise IOError('Pipeline cannot handle more than 6 replicates.')
seq_type = False if 'none' in IPs[
IPs.Condition == experiment]['File2'].tolist() else True
final_stage = 'align' if 'align' in IPs[
IPs.Condition == experiment]['Final Stage'].tolist() else 'all'
UMI_list = [
x.lower()
for x in IPs[IPs.Condition == experiment]['UMI'].unique().tolist()
]
if len(set(UMI_list)) > 1:
raise IOError(
'All samples must be UMI processed or not for each condition.')
UMI = True if UMI_list[0].lower() == 'yes' else False
try:
file_type = end_types[exp.sample_df[exp.sample_df.Condition ==
experiment]
['Scratch_File1'].tolist()[0][-4:]]
except KeyError:
output(
f"{exp.sample_df[exp.sample_df.Condition == experiment]['Scratch_File1'].tolist()[0]} not a valid file type for this pipeline.",
log_file=exp.log_file,
run_main=exp.run_main)
genome = IPs[IPs.Condition == experiment]['Genome'].unique().tolist()
if len(genome) > 1:
raise IOError(
'Cannot align to more than one genome per condition.')
chip_type = IPs[IPs.Condition ==
experiment]['ChIP Type'].unique().tolist()
if len(chip_type) > 1:
raise IOError(
'Cannot have more than one chip type (histone or TF) for a condition.'
)
chip_type = 'histone' if chip_type[0].lower() == 'histone' else 'tf'
json_file = {
'chip.pipeline_type': chip_type,
'chip.paired_end': seq_type,
'chip.genome_tsv': exp.genome_indicies['encode_tsv'][genome[0]],
'chip.bwa.mem_mb': 30000,
'chip.macs2_mem_mb': 30000,
'chip.peak_caller': 'macs2',
"chip.true_rep_only": False,
"chip.dup_marker": "picard",
"chip.mapq_thresh": 30,
"chip.regex_filter_reads": "chrM",
"chip.subsample_reads": 0,
"chip.ctl_subsample_reads": 0,
"chip.xcor_subsample_reads": 15000000,
"chip.keep_irregular_chr_in_bfilt_peak": False,
"chip.always_use_pooled_ctl": False,
"chip.ctl_depth_ratio": 1.2,
"chip.macs2_cap_num_peak": 500000,
"chip.pval_thresh": 0.01,
"chip.idr_thresh": 0.05,
"chip.bwa_cpu": 4,
"chip.bwa_mem_mb": 20000,
"chip.bwa_time_hr": 48,
"chip.filter_cpu": 2,
"chip.filter_mem_mb": 20000,
"chip.filter_time_hr": 24,
"chip.bam2ta_cpu": 2,
"chip.bam2ta_mem_mb": 10000,
"chip.bam2ta_time_hr": 6,
"chip.fingerprint_cpu": 2,
"chip.fingerprint_mem_mb": 12000,
"chip.fingerprint_time_hr": 6,
"chip.xcor_cpu": 2,
"chip.xcor_mem_mb": 16000,
"chip.xcor_time_hr": 24,
"chip.macs2_time_hr": 24,
"chip.spr_mem_mb": 16000
}
bams = []
ctl_bams = []
for rep, index in IP_sample_indicies:
sample = exp.sample_df.loc[index, 'Sample_Name']
input_sample = IPs.loc[index, 'Background_Name']
if file_type == 'fastq':
json_file[f'chip.fastqs_rep{rep}_R1'] = [
f'{exp.data_folder}{sample}_trim_R1.fastq.gz'
]
json_file[f'chip.ctl_fastqs_rep{rep}_R1'] = [
f'{exp.data_folder}{input_sample}_trim_R1.fastq.gz'
]
if seq_type:
json_file[f'chip.fastqs_rep{rep}_R2'] = [
f'{exp.data_folder}{sample}_trim_R2.fastq.gz'
]
json_file[f'chip.ctl_fastqs_rep{rep}_R2'] = [
f'{exp.data_folder}{input_sample}_trim_R2.fastq.gz'
]
else:
bams.append(f'{exp.data_folder}{sample}.bam')
ctl_bams.append(f'{exp.data_folder}{input_sample}.bam')
if file_type == 'bam':
json_file[f'chip.bams'] = bams
json_file[f'chip.ctl_bams'] = ctl_bams
json_file['chip.align_only'] = True if UMI & (file_type
== 'fastq') else False
json_file[
'chip.align_only'] = True if final_stage == 'align' else False
json_file['chip.no_dup_removal'] = True if UMI else False
json_file['chip.title'] = f'{experiment}_postUMI_dedup' if UMI & (
file_type == 'bam') else experiment
json_file[
"chip.description"] = f"Cromwell ENCODE3 {experiment}: {'paired-end' if seq_type else 'single-end'} {chip_type}."
encode_file = f'{exp_dir}{experiment}_ENCODE3.json'
with open(encode_file, 'w') as file:
json.dump(json_file, file, indent=4, sort_keys=True)
pythonpath = shutil.which('python')
miniconda = [x for x in pythonpath.split('/') if 'miniconda' in x]
cromwell_jar = re.sub(
r'{}/.*'.format(miniconda),
'{}/envs/chrome_chip/share/cromwell/cromwell.jar'.format(
miniconda), pythonpath)
jar = cromwell_jar if os.path.isfile(
cromwell_jar
) else '~/miniconda3/envs/chrome_chip/share/cromwell/cromwell.jar'
command_list = [
submission_prepend(source='encode-chip-seq-pipeline'),
f'cd {exp_dir}',
f'java -jar -Dconfig.file={exp.encode3_folder}backends/backend.conf -Dbackend.default=Local {jar} run {exp.encode3_folder}chip.wdl -i {encode_file}'
]
sent_job = send_job(command_list=command_list,
job_name=f"{experiment}_ENCODE3",
job_log_folder=exp.job_folder,
q='bigmem',
mem=35000,
log_file=exp.log_file,
project=exp.project,
cores=1,
run_main=exp.run_main)
exp.job_id.append(sent_job)
job_pending(sent_job, exp.log_file)
# Wait for jobs to finish
job_wait(exp.job_id, exp.log_file)
exp = encode_results(exp)
exp.tasks_complete.append('ENCODE3')
return exp
def encode3(exp):
if 'Stage' not in exp.tasks_complete:
output('Files not staged.\n', log_file=exp.log_file)
exp = stage(exp)
output('Running alignment and peak calling using ENCODE3 standards.',
log_file=exp.log_file,
run_main=exp.run_main)
output('ENCODE3 cromwell pipeline.',
log_file=exp.log_file,
run_main=exp.run_main)
out_dir = make_folder(f'{exp.scratch}ENCODE3/')
IPs = exp.IPs
end_types = {'q.gz': 'fastq', '.bam': 'bam'}
for experiment in IPs.Condition.unique().tolist():
exp_dir = make_folder(f'{out_dir}{experiment}/')
IP_sample_indicies = [(rep, index) for rep, index in enumerate(
IPs[IPs.Condition == experiment].index.tolist(), start=1)]
if len(IP_sample_indicies) > 6:
raise IOError('Pipeline cannot handle more than 6 replicates.')
seq_type = False if 'none' in IPs[
IPs.Condition == experiment]['File2'].tolist() else True
aligner = IPs[IPs.Condition == experiment]['Aligner'].unique().tolist()
if len(aligner) != 1:
raise IOError(
'All replicates must be aligned using the same aligner or not, which must be specified.'
)
else:
aligner = aligner[0]
peak_caller = IPs[IPs.Condition ==
experiment]['Peak Caller'].unique().tolist()
if len(peak_caller) != 1:
raise IOError(
'All replicates peaks must be called or not using the same peak calling strategy.'
)
else:
peak_caller = peak_caller[0]
UMI_list = [
x.lower()
for x in IPs[IPs.Condition == experiment]['UMI'].unique().tolist()
]
if len(set(UMI_list)) > 1:
raise IOError(
'All samples must be UMI processed or not for each condition.')
UMI = True if UMI_list[0] == 'yes' else False
try:
file_type = end_types[exp.sample_df[exp.sample_df.Condition ==
experiment]
['Scratch_File1'].tolist()[0][-4:]]
except KeyError:
output(
f"{exp.sample_df[exp.sample_df.Condition == experiment]['Scratch_File1'].tolist()[0]} not a valid file type for this pipeline.",
log_file=exp.log_file,
run_main=exp.run_main)
file_type = 'bam' if (UMI is True) & (
'UMI' in exp.tasks_complete) else file_type
genome = IPs[IPs.Condition == experiment]['Genome'].unique().tolist()
if len(genome) > 1:
raise IOError(
'Cannot align to more than one genome per condition.')
chip_type = IPs[IPs.Condition ==
experiment]['ChIP Type'].unique().tolist()
if len(chip_type) > 1:
raise IOError(
'Cannot have more than one chip type (histone or TF) for a condition.'
)
chip_type = 'histone' if chip_type[0].lower() == 'histone' else 'tf'
json_file = {
'chip.pipeline_type': chip_type,
'chip.paired_end': seq_type,
'chip.genome_tsv': exp.genome_indicies['encode_tsv'][genome[0]],
'chip.align_mem_mb': 30000,
"chip.true_rep_only": False,
"chip.dup_marker": "picard",
"chip.mapq_thresh": 30,
"chip.filter_chrs": ["chrM"],
"chip.subsample_reads": 0,
"chip.ctl_subsample_reads": 0,
"chip.xcor_subsample_reads": 15000000,
"chip.always_use_pooled_ctl": False,
"chip.ctl_depth_ratio": 1.2,
"chip.cap_num_peak_macs2": 500000,
"chip.pval_thresh": 0.01,
"chip.idr_thresh": 0.05,
"chip.align_cpu": 4,
"chip.align_time_hr": 48,
"chip.filter_cpu": 2,
"chip.filter_mem_mb": 20000,
"chip.filter_time_hr": 24,
"chip.bam2ta_cpu": 2,
"chip.bam2ta_mem_mb": 10000,
"chip.bam2ta_time_hr": 6,
"chip.jsd_cpu": 2,
"chip.jsd_mem_mb": 12000,
"chip.jsd_time_hr": 6,
"chip.xcor_cpu": 2,
"chip.xcor_mem_mb": 16000,
"chip.xcor_time_hr": 24,
"chip.align_time_hr": 24,
"chip.spr_mem_mb": 16000,
"chip.enable_count_signal_track": True,
}
if peak_caller == 'macs2':
json_file['chip.peak_caller'] = 'macs2'
if aligner != 'none':
json_file['chip.aligner'] = aligner
bams = []
ctl_bams = []
for rep, index in IP_sample_indicies:
sample = exp.sample_df.loc[index, 'Sample_Name']
input_sample = IPs.loc[index, 'Background_Name']
if file_type == 'fastq':
json_file[f'chip.fastqs_rep{rep}_R1'] = [
f'{exp.data_folder}{sample}_trim_R1.fastq.gz'
]
json_file[f'chip.ctl_fastqs_rep{rep}_R1'] = [
f'{exp.data_folder}{input_sample}_trim_R1.fastq.gz'
]
if seq_type:
json_file[f'chip.fastqs_rep{rep}_R2'] = [
f'{exp.data_folder}{sample}_trim_R2.fastq.gz'
]
json_file[f'chip.ctl_fastqs_rep{rep}_R2'] = [
f'{exp.data_folder}{input_sample}_trim_R2.fastq.gz'
]
else:
bams.append(f'{exp.data_folder}{sample}.bam')
ctl_bams.append(f'{exp.data_folder}{input_sample}.bam')
if file_type == 'bam':
json_file[f'chip.bams'] = bams
json_file[f'chip.ctl_bams'] = ctl_bams
json_file['chip.align_only'] = True if UMI & (file_type
== 'fastq') else False
json_file[
'chip.align_only'] = True if peak_caller == 'none' else json_file[
'chip.align_only']
json_file['chip.no_dup_removal'] = True if UMI else False
json_file['chip.title'] = f'{experiment}_postUMI_dedup' if UMI & (
file_type == 'bam') else experiment
json_file[
"chip.description"] = f"Cromwell ENCODE3 {experiment}: {'paired-end' if seq_type else 'single-end'} {chip_type}."
encode_file = f'{exp_dir}{experiment}_ENCODE3.json'
with open(encode_file, 'w') as file:
json.dump(json_file, file, indent=4, sort_keys=True)
pythonpath = shutil.which('python')
miniconda = [x for x in pythonpath.split('/') if 'miniconda' in x]
cromwell_jar = re.sub(
r'{}/.*'.format(miniconda),
'{}/envs/chrome_chip/share/cromwell/cromwell.jar'.format(
miniconda), pythonpath)
jar = cromwell_jar if os.path.isfile(
cromwell_jar
) else '~/miniconda3/envs/chrome_chip/share/cromwell/cromwell.jar'
command_list = [
submission_prepend(source='encode-chip-seq-pipeline'),
f'cd {exp_dir}',
f'java -jar -Dconfig.file={exp.encode3_folder}backends/backend.conf -Dbackend.default=Local {jar} run {exp.encode3_folder}chip.wdl -i {encode_file}'
]
sent_job = send_job(command_list=command_list,
job_name=f"{experiment}_ENCODE3",
job_log_folder=exp.job_folder,
q='bigmem',
mem=35000,
log_file=exp.log_file,
project=exp.project,
cores=1,
run_main=exp.run_main)
exp.job_id.append(sent_job)
job_pending(sent_job, exp.log_file)
# Wait for jobs to finish
job_wait(exp.job_id, exp.log_file)
# Check fraglength and resubmit with set 200 fraglen for macs2 if xcor error
for experiment in exp.IPs.Condition.unique().tolist():
rep_number = len(exp.IPs[exp.IPs.Condition == experiment])
frag_list = []
for rep in range(rep_number):
file = glob_check(
f'{exp.scratch}ENCODE3/{experiment}/cromwell-executions/chip/*/call-xcor/shard-{rep}/execution/*fraglen.txt'
)
with open(file, 'r') as f:
frag_list.append(f.read().split()[0])
if '-' in [x[0] for x in frag_list]:
output(
f'Xcor failed for {experiment}. Resubmitting with fragment length set to 200 for failed sample/s',
log_file=exp.log_file,
run_main=exp.run_main)
frag_list = [x if x[0] != '-' else '200' for x in frag_list]
exp_dir = f'{exp.scratch}ENCODE3/{experiment}/'
encode_file = f'{exp_dir}{experiment}_ENCODE3.json'
with open(encode_file, 'r') as file:
json_file = json.load(file)
json_file["chip.fraglen"] = frag_list
resubmit_file = f'{exp_dir}/{experiment}_ENCODE3_setfraglenth.json'
with open(resubmit_file, 'w') as file:
json.dump(json_file, file, indent=4, sort_keys=True)
pythonpath = shutil.which('python')
miniconda = [x for x in pythonpath.split('/') if 'miniconda' in x]
cromwell_jar = re.sub(
r'{}/.*'.format(miniconda),
'{}/envs/chrome_chip/share/cromwell/cromwell.jar'.format(
miniconda), pythonpath)
jar = cromwell_jar if os.path.isfile(
cromwell_jar
) else '~/miniconda3/envs/chrome_chip/share/cromwell/cromwell.jar'
command_list = [
submission_prepend(source='encode-chip-seq-pipeline'),
f'cd {exp_dir}',
f'java -jar -Dconfig.file={exp.encode3_folder}backends/backend.conf -Dbackend.default=Local {jar} run {exp.encode3_folder}chip.wdl -i {resubmit_file}'
]
sent_job = send_job(command_list=command_list,
job_name=f"{experiment}_ENCODE3_resubmission",
job_log_folder=exp.job_folder,
q='bigmem',
mem=35000,
log_file=exp.log_file,
project=exp.project,
cores=1,
run_main=exp.run_main)
exp.job_id.append(sent_job)
job_pending(sent_job, exp.log_file)
# Wait for jobs to finish
job_wait(exp.job_id, exp.log_file)
exp = encode_results(exp)
exp.tasks_complete.append('ENCODE3')
return exp
cmd = f'python chrome_chip -f {args.experimental_file}'
if args.template_notebook:
cmd += f' -t {args.template_notebook}'
if args.out_notebook:
cmd += f' -o {args.out_notebook}'
submission_header = [submission_prepend(cmd)]
job_name = args.experimental_file.split('.')[0]
send_job(command_list=submission_header,
job_name=job_name,
job_log_folder=f'{os.getcwd()}/',
q='general',
mem=3000,
log_file=f'bsub_{job_name}.log',
project=args.project,
cores=1,
submit=True,
run_main=False
)
else:
if args.template_notebook:
if os.path.isfile(args.template_notebook) is False:
raise IOError(f'Location of template notebook not found. Use -t option.')
else:
import papermill as pm
out_notebook = args.out_notebook if args.out_notebook else args.experimental_file.replace('yml', 'ipynb')
pm.execute_notebook(args.template_notebook, out_notebook, parameters=dict(yaml_file=args.experimental_file), log_output=True, report_mode=True)
