rule not picked up by snakemake - snakemake

I'm starting with snakemake. I managed to define some rules which I can run indepently, but not in a workflow. Maybe the issue is that they have unrelated inputs and outputs.
My current workflow is like this:
configfile: './config.yaml'
rule all:
input: dynamic("task/{job}/taskOutput.tab")
rule split_input:
input: "input_fasta/snp.fa"
output: dynamic("task/{job}/taskInput.fa")
shell:
"rm -Rf tasktmp task; \
mkdir tasktmp task; \
split -l 200 -d {input} ./tasktmp/; \
ls tasktmp | awk '{{print \"mkdir task/\"$0}}' | sh; \
ls tasktmp | awk '{{print \"mv ./tasktmp/\"$0\" ./task/\"$0\"/taskInput.fa\"}}' | sh"
rule task:
input: "task/{job}/taskInput.fa"
output: "task/{job}/taskOutput.tab"
shell: "cp {input} {output}"
rule make_parameter_file:
output:
"par/parameters.txt
shell:
"rm -Rf par;mkdir par; \
echo \"\
minimumFlankLength=5\n\
maximumFlankLength=200\n\
alignmentLengthDifference=2\
allowedMismatch=4\n\
allowedProxyMismatch=2\n\
allowedIndel=3\n\
ambiguitiesAsMatch=1\n\" \
> par/parameters.txt"
rule build_target:
input:
"./my_target"
output:
touch("build_target.done")
shell:
"build_target -template format_nt -source {input} -target my_target"
If I call this as such:
snakemake -p -s snakefile
The first three rules are being executed, the others not.
I can run the last rule by specifying it as an argument.
snakemake -p -s snakefile build_target
But I don't see how I can run all.
Thanks a lot for any suggestion on how to solve this.

By default snakemake executes only the first rule of a snakefile. Here it is rule all. In order to produce rule all's input dynamic("task/{job}/taskOutput.tab"), it needs to run the following two rules task and split_input, and so it does.
If you want the other rules to be run as well, you should put their output in rule all, eg.:
rule all:
input:
dynamic("task/{job}/taskOutput.tab"),
"par/parameters.txt",
"build_target.done"

Related

Snakemake Megahit output issue

A few days ago I started using Snakemake for the first time. I am having an issue when I am trying to run the megahit rule in my pipeline.
It gives me the following error "Outputs of incorrect type (directories when expecting files or vice versa). Output directories must be flagged with directory(). ......"
So initially it runs and then crashes with the above error. I implemented the solution with the directory() option in my pipeline but I think its not a good practice since, for various reasons, you can loose files without even knowing it.
Is there a way to run the rule without using the directory() ?
I would appreciate any help on the issue!
Thanking you in advance
sra = []
with open("run_ids") as f:
for line in f:
sra.append(line.strip())
rule all:
input:
expand("raw_reads/{sample}/{sample}.fastq", sample=sra),
expand("trimmo/{sample}/{sample}.trimmed.fastq", sample=sra),
expand("megahit/{sample}/final.contigs.fa", sample=sra)
rule download:
output:
"raw_reads/{sample}/{sample}.fastq"
params:
"--split-spot --skip-technical"
log:
"logs/fasterq-dump/{sample}.log"
benchmark:
"benchmarks/fastqdump/{sample}.fasterq-dump.benchmark.txt"
threads: 8
shell:
"""
fasterq-dump {params} --outdir /home/raw_reads/{wildcards.sample} {wildcards.sample} -e {threads}
"""
rule trim:
input:
"raw_reads/{sample}/{sample}.fastq"
output:
"trimmo/{sample}/{sample}.trimmed.fastq"
params:
"HEADCROP:15 TRAILING:3 SLIDINGWINDOW:4:15 MINLEN:36"
log:
"logs/trimmo/{sample}.log"
benchmark:
"benchmarks/trimmo/{sample}.trimmo.benchmark.txt"
threads: 6
shell:
"""
trimmomatic SE -phred33 -threads {threads} {input} trimmo/{wildcards.sample}/{wildcards.sample}.trimmed.fastq {params}
"""
rule megahit:
input:
"trimmo/{sample}/{sample}.trimmed.fastq"
output:
"megahit/{sample}/final.contigs.fa"
params:
"-m 0.7 -t"
log:
"logs/megahit/{sample}.log"
benchmark:
"benchmarks/megahit/{sample}.megahit.benchmark.txt"
threads: 10
shell:
"""
megahit -r {input} -o {output} -t {threads}
"""
IMHO it is a bad design of the megahit software that it takes a directory as a parameter and outputs into a file in this directory with a hardcoded name. Flagging the filename with directory() doesn't solve the issue, as in this case what you expect to be a file with the .fa extension megahit treats as a directory. The rest of the pipeline is broken in this case.
But this issue can be solved in Snakemake like that:
rule megahit:
input:
"trimmo/{sample}/{sample}.trimmed.fastq"
output:
"megahit/{sample}/final.contigs.fa"
# ...
shell:
"""
megahit -r {input} -o megahit/{wildcards.sample} -t {threads}
"""
A better design of the megahit rule would look as follows:
rule megahit:
input:
"trimmo/{sample}/{sample}.trimmed.fastq"
output:
out_dir = directory("megahit/{sample}/"),
fasta = "megahit/{sample}/final.contigs.fa"
log:
"logs/megahit/{sample}.log"
benchmark:
"benchmarks/megahit/{sample}.megahit.benchmark.txt"
threads:
10
shell:
"megahit -r {input} -f -o {output.out_dir} -t {threads}"
This guarantees that the output directory is removed upon failure, while the -f argument to megahit tells it to ignore the fact that the output folder exists (it is created by Snakemake automatically because one of the outputs is a file inside it: final.contigs.fa).
BTW, the -m (--memory) parameter is best implemented as a resource. The only problem though is that snakemake's default resource, mem_mb is in megabytes. One workaround would be as follows:
resources:
mem_mb = mem_mb_limit_for_megahit # could be a fraction of a global constant
params:
mem_bytes = lambda w, resources: round(resources.mem_mb * 1e6)
shell:
"megahit ... -m {params.mem_bytes}"

snakemake - replacing command line parameters with wildcards by cluster profile

I am writing a snakemake pipeline to eventually identify corona virus variants.
Below is a minimal example with three steps:
LOGDIR = '/path/to/logDir'
barcodes = ['barcode49', 'barcode50', 'barcode51']
rule all:
input:
expand([
# guppyplex
"out/guppyplex/{barcode}/{barcode}.fastq",
# catFasta
"out/catFasta/cat_consensus.fasta",
], barcode = barcodes)
rule guppyplex:
input:
FQ = f"fastq/{{barcode}}" # FASTQ_PATH is parsed from config.yaml
output:
"out/guppyplex/{barcode}/{barcode}.fastq"
shell:
"touch {output}" # variables in CAPITALS are parsed from config.yaml
rule minion:
input:
INFQ = rules.guppyplex.output,
FAST5 = f"fasta/{{barcode}}"
params:
OUTDIR = "out/nanopolish/{barcode}"
output:
"out/nanopolish/{barcode}/{barcode}.consensus.fasta"
shell:
"""
touch {output} && echo {wildcards.barcode} > {output}
"""
rule catFasta:
input:
expand("out/nanopolish/{barcode}/{barcode}.consensus.fasta", barcode = barcodes)
output:
"out/catFasta/cat_consensus.fasta"
shell:
"cat {input} > {output}"
If I run the snakemake locally by calling snakemake -p --cores 1 all everything works. Yet my ultimate goal is to use qsub to run the jobs on a cluster. I also want the stderr and stdout from qsub to have meaningful names, which include wildcards and the rule names for each job.
However, if I call snakemake with
snakemake -p --cluster "qsub -q onlybngs05b -e {LOGDIR} -o {LOGDIR} -j y" -j 5 --jobname "{wildcards.barcode}.{rule}.{jobid}" all
I will get the following error:
AttributeError: 'Wildcards' object has no attribute 'barcode'
I have recently read the snakemake documentation where it appears that I could replace the command line parameters (--cluster "qsub -q onlybngs05b -e {LOGDIR} -o {LOGDIR} -j y" -j 5 --jobname "{wildcards.barcode}.{rule}.{jobid}") by a yaml file. Although the documentation is not all that clear to me.
I have created a config.yaml file at /home/user/.config/snakemake which looks like so:
cluster: 'qsub'
q: 'onlybngs05b'
e: '/home/ngs/tempOutSnakemake'
o: '/home/ngs/tempOutSnakemake'
j: 5
jobname: "{wildcards.barcode}.{rule}.{jobid}
But then it appears that snakemake is not properly parsing the config.yaml. I am getting
snakemake: error: ambiguous option: --o=/home/ngs/tempOutSnakemake could match --omit-from, --output-wait, --overwrite-shellcmd
I also tried to replace o in the config file by stdout (kind of the long version of the parameter (-h vs --help for several programs), though it does not work.
Therefore my question is how I can replace the command line parameters --cluster "qsub -q onlybngs05b -e {LOGDIR} -o {LOGDIR} -j y" -j 5 --jobname "{wildcards.barcode}.{rule}.{jobid}" by a config.yaml file that accepts wildcards?
I think the problem is that rule catFasta doesn't contain the wildcard barcode. If you think about it, what job name would you expect in {wildcards.barcode}.{rule}.{jobid}?
Maybe a solution could be to add to each rule a jobname parameter that could be {barcode} for guppyplex and minion and 'all_barcodes' for catFasta. Then use --jobname "{params.jobname}.{rule}.{jobid}"

Command not found error in snakemake pipeline despite the package existing in the conda environment

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)

Nothing to be done for running snakefile

My code is like this, I have set the rule all:
rule all:
input:
expand("data/sam/{sample}.sam", sample=SAMPLE_NAMES)
rule trimmomatic:
input:
"data/samples/{sample}.fastq"
output:
"data/samples/{sample}.clean.fastq"
shell:
"trimmomatic SE -threads 5 -phred33 -trimlog trim.log {input} {output} LEADING:20 TRAILING:20 MINLEN:16"
rule hisat2:
input:
fa="data/genome.fa",
fastq="data/samples/{sample}.clean.fastq"
output:
"data/sam/{sample}.sam"
shell:
"hisat2-build {input.fa} ./index/geneindex | hisat2 -x - -q samples/{inout.fastq} -S {output}"
but it still shows that :
Nothing to be done.
I have try to find the way out. but useless.
help!

Snakemake: remove output file

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}"""