I show below a pseudocode version of my snakefile. Snakemake rule A creates the input files for Snakemake rule B2 and I would like to run Snakemake rules B1 and B2 at the same time but am not having success. I can run this snakefile successfully on very small data without a problem (although the Snakemake rules B1 and B2 do not run in parallel) but once I give it larger data it fails to create the output for Snakemake rule B1. The commands between Snakemake rule B1 and B2 use the same program but have different arguments and input files so I didn't think they should be in the same rule.
rule all:
input: file_A_out, file_B1_out, file_B2_out, file_C_out
rule A:
input: file_A_in
output: file_A_out
log: file_A_log
shell: 'progA {input} --output {output}'
rule B1:
input: file_B1_in
output: file_B1_out
group: 'groupB'
log: file_B1_log
shell: 'progB {input} -x 100 -o {output}'
rule B2:
input: file_A_out
output: file_B2_out
group: 'groupB'
log: file_B2_log
shell: 'progB {input} -x 1 --y -o {output}'
rule C:
input: file_B1_out, file_B2_out
output: file_C_out
log: file_C_log
shell: 'progC {input[0]} {input[1]} -o {output}'
I thought using group to group the rules would indicate to Snakemake that the two rules can be ran at once. To execute snakemake I run nohup snakemake --cores 16 > log.txt 2>&1 & however, it only successfully runs rule B2 while the output of rule B1 is deemed corrupted. I have seen solutions on running one rule in parallel but what about running different rules in parallel?
Error in rule B1:
jobid: 2
input: 'file_B1_in'
output: 'file_B1_out'
log: 'file_B1_log'
(check log file(s) for error details)
shell: 'progB {input} -x 100 -o {output}'
(one of the commands exited with non-zero exit code; note that snakemake uses bash strict mode!)
Removing output files of failed job B1 since they might be corrupted:
file_B1_out
Shutting down, this might take some time.
Exiting because a job execution failed. Look above for error message
The snakefile below runs rules A, B1, and B2 in parallel then runs rule C, as expected. Maybe there is something you are not showing us?
# Make dummy input files
touch file_A_in file_B1_in
# Run pipeline
snakemake -p -j 10
The snakefile:
rule all:
input: 'file_A_out', 'file_B1_out', 'file_B2_out', 'file_C_out'
rule A:
input: 'file_A_in'
output: 'file_A_out'
shell: 'sleep 10; echo {input} > {output}'
rule B1:
input: 'file_B1_in'
output: 'file_B1_out'
shell: 'sleep 10; echo {input} > {output}'
rule B2:
input: 'file_A_in'
output: 'file_B2_out'
shell: 'sleep 10; echo {input} > {output}'
rule C:
input: 'file_B1_out', 'file_B2_out'
output: 'file_C_out'
shell: 'sleep 10; echo {input[0]} {input[1]} > {output}'
Related
This is a very strange problem.
When my {input} specified in the rule section is a list of <200 files, snakemake worked all right.
But when {input} has more than 500 files, snakemake just quitted with messages (one of the commands exited with non-zero exit code; note that snakemake uses bash strict mode!). The complete log did not provide any error messages.
For the log, please see: https://github.com/snakemake/snakemake/files/5285271/2020-09-25T151835.613199.snakemake.log
The rule that worked is (NOTE the input is capped to 200 files):
rule combine_fastq:
input:
lambda wildcards: samples.loc[(wildcards.sample), ["fq"]].dropna()[0].split(',')[:200]
output:
"combined.fastq/{sample}.fastq.gz"
group: "minion_assemble"
shell:
"""
echo {input} > {output}
"""
The rule that failed is:
rule combine_fastq:
input:
lambda wildcards: samples.loc[(wildcards.sample), ["fq"]].dropna()[0].split(',')
output:
"combined.fastq/{sample}.fastq.gz"
group: "minion_assemble"
shell:
"""
echo {input} > {output}
"""
My question is also posted in GitHub: https://github.com/snakemake/snakemake/issues/643.
I second Maarten's answer, with that many files you are running up against a shell limit; snakemake is just doing a poor job helping you identify the problem.
Based on the issue you reference, it seems like you are using cat to combine all of your files. Maybe following the answer here would help:
rule combine_fastq_list:
input:
lambda wildcards: samples.loc[(wildcards.sample), ["fq"]].dropna()[0].split(',')
output:
temp("{sample}.tmp.list")
group: "minion_assemble"
script:
with open(output[0]) as out:
out.write('\n'.join(input))
rule combine_fastq:
input:
temp("{sample}.tmp.list")
output:
'combined.fastq/{sample}.fastq.gz'
group: "minion_assemble"
shell:
'cat {input} | ' # this is reading the list of files from the file
'xargs zcat -f | '
'...'
Hope it gets you on the right track.
edit
The first option executes your command separately for each input file. A different option that executes the command once for the whole list of input is:
rule combine_fastq:
...
shell:
"""
command $(< {input}) ...
"""
For those landing here with similar questions (like Snakemake expand function alternative), snakemake 6 can handle long command lines. The following test fails on snakemake < 6 but succeeds on 6.0.0 on my Ubuntu machine:
rule all:
input:
'output.txt',
rule one:
output:
'output.txt',
params:
x= list(range(0, 1000000))
shell:
r"""
echo {params.x} > {output}
"""
I am using an assembly pipeline called Canu inside my snakemake pipeline, but when it comes to the rule calling Canu, snakemake exits witht he MissingOutputException error as the pipeline submits multiple jobs to the cluster itself so it seems snakemake expects the output after the first job has finished. Is there a way to avoid this? I know I could use a very long --latency-wait option but this is not very optimal.
snakefile code:
#!/miniconda/bin/python
workdir: config["path_to_files"]
wildcard_constraints:
separator = config["separator"],
sample = '|' .join(config["samples"]),
rule all:
input:
expand("assembly-stats/{sample}_stats.txt", sample = config["samples"])
rule short_reads_QC:
input:
f"short_reads/{{sample}}_short{config['separator']}*.fq.gz"
output:
"fastQC-reports/{sample}.html"
conda:
"/home/lamma/env-export/hybrid_assembly.yaml"
shell:
"""
mkdir fastqc-reports
fastqc -o fastqc-reports {input}
"""
rule quallity_trimming:
input:
forward = f"short_reads/{{sample}}_short{config['separator']}1.fq.gz",
reverse = f"short_reads/{{sample}}_short{config['separator']}2.fq.gz",
output:
forward = "cleaned_short-reads/{sample}_short_1-clean.fastq",
reverse = "cleaned_short-reads/{sample}_short_2-clean.fastq"
conda:
"/home/lamma/env-export/hybrid_assembly.yaml"
shell:
"bbduk.sh -Xmx1g in1={input.forward} in2={input.reverse} out1={output.forward} out2={output.reverse} qtrim=rl trimq=10"
rule long_read_assembly:
input:
"long_reads/{sample}_long.fastq.gz"
output:
"canu-outputs/{sample}.subreads.contigs.fasta"
conda:
"/home/lamma/env-export/hybrid_assembly.yaml"
shell:
"canu -p {wildcards.sample} -d canu-outputs genomeSize=8m -pacbio-raw {input}"
rule short_read_alignment:
input:
short_read_fwd = "cleaned_short-reads/{sample}_short_1-clean.fastq",
short_read_rvs = "cleaned_short-reads/{sample}_short_2-clean.fastq",
reference = "canu-outputs/{sample}.subreads.contigs.fasta"
output:
"bwa-output/{sample}_short.bam"
conda:
"/home/lamma/env-export/hybrid_assembly.yaml"
shell:
"bwa mem {input.reference} {input.short_read_fwd} {input.short_read_rvs} | samtools view -S -b > {output}"
rule indexing_and_sorting:
input:
"bwa-output/{sample}_short.bam"
output:
"bwa-output/{sample}_short_sorted.bam"
conda:
"/home/lamma/env-export/hybrid_assembly.yaml"
shell:
"samtools sort {input} > {output}"
rule polishing:
input:
bam_files = "bwa-output/{sample}_short_sorted.bam",
long_assembly = "canu-outputs/{sample}.subreads.contigs.fasta"
output:
"pilon-output/{sample}-improved.fasta"
conda:
"/home/lamma/env-export/hybrid_assembly.yaml"
shell:
"pilon --genome {input.long_assembly} --frags {input.bam_files} --output {output} --outdir pilon-output"
rule assembly_stats:
input:
"pilon-output/{sample}-improved.fasta"
output:
"assembly-stats/{sample}_stats.txt"
conda:
"/home/lamma/env-export/hybrid_assembly.yaml"
shell:
"stats.sh in={input} gc=assembly-stats/{wildcards.sample}/{wildcards.sample}_gc.csv gchist=assembly-stats/{wildcards.sample}/{wildcards.sample}_gchist.csv shist=assembly-stats/{wildcards.sample}/{wildcards.sample}_shist.csv > assembly-stats/{wildcards.sample}/{wildcards.sample}_stats.txt"
The exact error:
Waiting at most 60 seconds for missing files.
MissingOutputException in line 43 of /faststorage/home/lamma/scripts/hybrid_assembly/bacterial-hybrid-assembly.smk:
Missing files after 60 seconds:
canu-outputs/F19FTSEUHT1027.PSU4_ISF1A.subreads.contigs.fasta
This might be due to filesystem latency. If that is the case, consider to increase the wait time with --latency-wait.
Shutting down, this might take some time.
Exiting because a job execution failed. Look above for error message
The snakemake command being used:
snakemake --latency-wait 60 --rerun-incomplete --keep-going --jobs 99 --cluster-status 'python /home/lamma/faststorage/scripts/slurm-status.py' --cluster 'sbatch -t {cluster.time} --mem={cluster.mem} --cpus-per-task={cluster.c} --error={cluster.error} --job-name={cluster.name} --output={cluster.output}' --cluster-config bacterial-hybrid-assembly-config.json --configfile yaml-config-files/test_experiment3.yaml --use-conda --snakefile bacterial-hybrid-assembly.smk
I surmise that canu is giving you canu-outputs/{sample}.contigs.fasta not canu-outputs/{sample}.subreads.contigs.fasta. If so edit the canu commad to be
canu -p {wildcards.sample}.subreads ...
(By the way, I don't think #!/miniconda/bin/python is necessary).
I am getting the following error in the snakemake pipeline:
Building DAG of jobs...
Using shell: /usr/bin/bash
Provided cores: 16
Rules claiming more threads will be scaled down.
Job counts:
count jobs
1 long_read_assembly
1
[Wed Jan 15 11:35:18 2020]
rule long_read_assembly:
input: long_reads/F19FTSEUHT1027.PSU4_ISF1A_long.fastq.gz
output: canu-outputs/F19FTSEUHT1027.PSU4_ISF1A.subreads.contigs.fasta
jobid: 0
wildcards: sample=F19FTSEUHT1027.PSU4_ISF1A
/usr/bin/bash: canu: command not found
[Wed Jan 15 11:35:18 2020]
Error in rule long_read_assembly:
jobid: 0
output: canu-outputs/F19FTSEUHT1027.PSU4_ISF1A.subreads.contigs.fasta
shell:
canu -p F19FTSEUHT1027.PSU4_ISF1A -d canu-outputs genomeSize=8m -pacbio-raw long_reads/F19FTSEUHT1027.PSU4_ISF1A_long.fastq.gz
(one of the commands exited with non-zero exit code; note that snakemake uses bash strict mode!)
Shutting down, this might take some time.
Exiting because a job execution failed. Look above for error message
I assume it is meaning that the command canu can not be found. But the Canu package does exist inside the conda environment:
(hybrid_assembly) [lamma#fe1 Assembly]$ conda list | grep canu
canu 1.9 he1b5a44_0 bioconda
The snakefile looks like this:
workdir: config["path_to_files"]
wildcard_constraints:
separator = config["separator"],
sample = '|' .join(config["samples"]),
rule all:
input:
expand("assembly-stats/{sample}_stats.txt", sample = config["samples"])
rule short_reads_QC:
input:
f"short_reads/{{sample}}_short{config['separator']}*.fq.gz"
output:
"fastQC-reports/{sample}.html"
conda:
"/home/lamma/env-export/hybrid_assembly.yaml"
shell:
"""
mkdir fastqc-reports
fastqc -o fastqc-reports {input}
"""
rule quallity_trimming:
input:
forward = f"short_reads/{{sample}}_short{config['separator']}1.fq.gz",
reverse = f"short_reads/{{sample}}_short{config['separator']}2.fq.gz",
output:
forward = "cleaned_short-reads/{sample}_short_1-clean.fastq",
reverse = "cleaned_short-reads/{sample}_short_2-clean.fastq"
conda:
"/home/lamma/env-export/hybrid_assembly.yaml"
shell:
"bbduk.sh -Xmx1g in1={input.forward} in2={input.reverse} out1={output.forward} out2={output.reverse} qtrim=rl trimq=10"
rule long_read_assembly:
input:
"long_reads/{sample}_long.fastq.gz"
output:
"canu-outputs/{sample}.subreads.contigs.fasta"
conda:
"/home/lamma/env-export/hybrid_assembly.yaml"
shell:
"canu -p {wildcards.sample} -d canu-outputs genomeSize=8m -pacbio-raw {input}"
rule short_read_alignment:
input:
short_read_fwd = "cleaned_short-reads/{sample}_short_1-clean.fastq",
short_read_rvs = "cleaned_short-reads/{sample}_short_2-clean.fastq",
reference = "canu-outputs/{sample}.subreads.contigs.fasta"
output:
"bwa-output/{sample}_short.bam"
conda:
"/home/lamma/env-export/hybrid_assembly.yaml"
shell:
"bwa mem {input.reference} {input.short_read_fwd} {input.short_read_rvs} | samtools view -S -b > {output}"
rule indexing_and_sorting:
input:
"bwa-output/{sample}_short.bam"
output:
"bwa-output/{sample}_short_sorted.bam"
conda:
"/home/lamma/env-export/hybrid_assembly.yaml"
shell:
"samtools sort {input} > {output}"
rule polishing:
input:
bam_files = "bwa-output/{sample}_short_sorted.bam",
long_assembly = "canu-outputs/{sample}.subreads.contigs.fasta"
output:
"pilon-output/{sample}-improved.fasta"
conda:
"/home/lamma/env-export/hybrid_assembly.yaml"
shell:
"pilon --genome {input.long_assembly} --frags {input.bam_files} --output {output} --outdir pilon-output"
rule assembly_stats:
input:
"pilon-output/{sample}-improved.fasta"
output:
"assembly-stats/{sample}_stats.txt"
conda:
"/home/lamma/env-export/hybrid_assembly.yaml"
shell:
"stats.sh in={input} gc=assembly-stats/{wildcards.sample}/{wildcards.sample}_gc.csv gchist=assembly-stats/{wildcards.sample}/{wildcards.sample}_gchist.csv shist=assembly-stats/{wildcards.sample}/{wildcards.sample}_shist.csv > assembly-stats/{wildcards.sample}/{wildcards.sample}_stats.txt"
The rule calling canu has the correct syntax as far as I am awear so I am not sure what is causing this error.
Edit:
Adding the snakemake command
snakemake --latency-wait 60 --rerun-incomplete --keep-going --jobs 99 --cluster-status 'python /home/lamma/faststorage/scripts/slurm-status.py' --cluster 'sbatch -t {cluster.time} --mem={cluster.mem} --cpus-per-task={cluster.c} --error={cluster.error} --job-name={cluster.name} --output={cluster.output} --wait --parsable' --cluster-config bacterial-hybrid-assembly-config.json --configfile yaml-config-files/test_experiment3.yaml --snakefile bacterial-hybrid-assembly.smk
When running a snakemake workflow, if certain rules are to be ran within a rule-specific conda environment, the command line call should be of the form
snakemake [... various options ...] --use-conda [--conda-prefix <some-directory>]
If you don't tell snakemake to use conda, all the conda: <some_path> entries in your rules are ignored, and the rules are run in whatever environment is currently activated.
The --conda-prefix <dir> is optional, but tells snakemake where to find the installed environment (if you don't specify this, a conda env will be installed within the .snakemake folder, meaning that the .snakemake folder can get pretty huge and that the .snakemake folders for multiple projects may contain a lot of duplicated conda stuff)
So I have an issue when I run other programs after I ran flye in my snakemake pipeline. This is because the output from flye is a directory. My rules are as followd:
samples, = glob_wildcards("data/samples/{sample}.fastq")
rule all:
input:
[f"assembled/" for sample in samples],
[f"nanopolish/draft.fa" for sample in samples],
[f"nanopolish/reads.sorted.bam" for sample in samples],
[f"nanopolish/reads.indexed.sorted.bam" for sample in samples]
rule fly:
input:
"unzipped/read.fastq"
output:
directory("assembled/")
conda:
"envs/flye.yaml"
shell:
"flye --nano-corr {input} --genome-size 5m --out-dir {output}"
rule bwa:
input:
"assembled/assembly.fasta"
output:
"nanopolish/draft.fa"
conda:
"nanopolish.yaml"
shell:
"bwa index {input} {output}"
rule nanopolish:
input:
"nanopolish/draft.fa",
"zipped/zipped.gz"
output:
"nanopolish/reads.sorted.bam"
conda:
"nanopolish.yaml"
shell:
"bwa mem -x ont2d -t 8 {input} | samtools sort -o {output}"
there are a few steps before this but they work just fine. when I run this it gives the following error:
ChildIOException:
File/directory is a child to another output:
/home/fronglesquad/snakemake_poging_1/assembled
/home/fronglesquad/snakemake_poging_1/assembled/assembly.fasta
I have googled the error. All I could find there that its because snakemake doesnt work well with output directorys. But this tool needs a output directory to work. Does anyone know how to bypass this?
(I think) The problem lies somewhere else in your code.
You have defined two rules, the first that outputs directory assembled, the second that outputs assembled/assembly.fasta. Since the output of the second rule is always at least the directory assembled, Snakemake complains. You can solve it by using the directory as input:
rule second:
input:
"assembled"
output:
...
shell:
cat {input}/assembly.fasta > {output}
I don't see how to use a Snakemake rule to remove a Snakemake output file that has become useless.
In concrete terms, I have a rule bwa_mem_sam that creates a file named {sample}.sam.
I have this other rule, bwa_mem_bam that creates a file named {sample.bam}.
Has the two files contain the same information in different formats, I'd like to remove the first one cannot succeed doing this.
Any help would be very much appreciated.
Ben.
rule bwa_mem_map:
input:
sam="{sample}.sam",
bam="{sample}.bam"
shell:
"rm {input.sam}"
# Convert SAM to BAM.
rule bwa_mem_map_bam:
input:
rules.sam_to_bam.output
# Use bwa mem to map reads on a reference genome.
rule bwa_mem_map_sam:
input:
reference=reference_genome(),
index=reference_genome_index(),
fastq=lambda wildcards: config["units"][SAMPLE_TO_UNIT[wildcards.sample]],
output:
"mapping/{sample}.sam"
threads: 12
log:
"mapping/{sample}.log"
shell:
"{BWA} mem -t {threads} {input.reference} {input.fastq} > {output} 2> {log} "\
"|| (rc=$?; cat {log}; exit $rc;)"
rule sam_to_bam:
input:
"{prefix}.sam"
output:
"{prefix}.bam"
threads: 8
shell:
"{SAMTOOLS} view --threads {threads} -b {input} > {output}"
You don't need a rule to remove you sam files. Just mark the ouput sam file in "bwa_mem_map_sam" rule as temporary:
rule bwa_mem_map_sam:
input:
reference=reference_genome(),
index=reference_genome_index(),
fastq=lambda wildcards: config["units"][SAMPLE_TO_UNIT[wildcards.sample]],
output:
temp("mapping/{sample}.sam")
threads: 12
log:
"mapping/{sample}.log"
shell:
"{BWA} mem -t {threads} {input.reference} {input.fastq} > {output} 2> {log} "\
"|| (rc=$?; cat {log}; exit $rc;)"
as soon as a temp file is not needed anymore (ie: not used as input in any other rule), it will be removed by snakemake.
EDIT AFTER COMMENT:
If I understand correctly, your statement "if the user asks for a sam..." means the sam file is put in the target rule. If this is the case, then as long as the input of the target rule contains the sam file, the file won't be deleted (I guess). If the bam file is put in the target rule (and not the sam), then it will be deleted.
The other way is this:
rule bwa_mem_map:
input:
sam="{sample}.sam",
bam="{sample}.bam"
output:
touch("{sample}_samErased.txt")
shell:
"rm {input.sam}"
and ask for "{sample}_samErased.txt" in the target rule.
Based on the comments above, you want to ask the user if he wants a sam or bam output.
You could use this as a config argument:
snakemake --config output_format=sam
Then you use this kind Snakefile:
samples = ['A','B']
rule all:
input:
expand('{sample}.mapped.{output_format}', sample=samples, output_format=config['output_format'])
rule bwa:
input: '{sample}.fastq'
output: temp('{sample}.mapped.sam')
shell:
"""touch {output}"""
rule sam_to_bam:
input: '{sample}.mapped.sam'
output: '{sample}.mapped.bam'
shell:
"""touch {output}"""