Hello, I'm a graduated student studying bioinformatics in South Korea.
I'm trying to do detect extrachromosomal DNA (ecDNA) in nanopore long-read targeted sequencing data.
I have tried two times of that workflow, but had same error in my job.
Workflow defines that rule get_genome is eligible for caching between workflows (use the --cache argument to enable this).
Workflow defines that rule genome_faidx is eligible for caching between workflows (use the --cache argument to enable this).
Workflow defines that rule minimap2_index is eligible for caching between workflows (use the --cache argument to enable this).
Workflow defines that rule download_regulatory_annotation is eligible for caching between workflows (use the --cache argument to enable this).
Workflow defines that rule download_repeatmasker_annotation is eligible for caching between workflows (use the --cache argument to enable this).
Workflow defines that rule download_gene_annotation is eligible for caching between workflows (use the --cache argument to enable this).
Assuming unrestricted shared filesystem usage.
Building DAG of jobs...
Your conda installation is not configured to use strict channel priorities. This is however crucial for having robust and correct environments (for details, see https://conda-forge.org/docs/user/tipsandtricks.html). Please consider to configure strict priorities by executing 'conda config --set channel_priority strict'.
Creating conda environment https://raw.githubusercontent.com/snakemake-workflows/cyrcular-calling/v2.1.0/workflow/envs/python.yaml...
Downloading and installing remote packages.
Environment for https://raw.githubusercontent.com/snakemake-workflows/cyrcular-calling/v2.1.0/workflow/envs/python.yaml created (location: .snakemake/conda/f73c94fe37d96f51957feb7810eab8cf_)
Creating conda environment https://github.com/snakemake/snakemake-wrappers/raw/v1.25.0/bio/bcftools/concat/environment.yaml...
Downloading and installing remote packages.
Environment for https://github.com/snakemake/snakemake-wrappers/raw/v1.25.0/bio/bcftools/concat/environment.yaml created (location: .snakemake/conda/cf33438f5c7c9884abf85d29285efe04_)
Creating conda environment https://raw.githubusercontent.com/snakemake-workflows/cyrcular-calling/v2.1.0/workflow/envs/varlociraptor.yaml...
Downloading and installing remote packages.
Environment for https://raw.githubusercontent.com/snakemake-workflows/cyrcular-calling/v2.1.0/workflow/envs/varlociraptor.yaml created (location: .snakemake/conda/2eef35d24a59f83df9961f5f70ab7240_)
Creating conda environment https://raw.githubusercontent.com/snakemake-workflows/cyrcular-calling/v2.1.0/workflow/envs/graphviz.yaml...
Downloading and installing remote packages.
Environment for https://raw.githubusercontent.com/snakemake-workflows/cyrcular-calling/v2.1.0/workflow/envs/graphviz.yaml created (location: .snakemake/conda/359230e4a02075ecdb5b70d4ee000c2b_)
Creating conda environment https://raw.githubusercontent.com/snakemake-workflows/cyrcular-calling/v2.1.0/workflow/envs/bcftools.yaml...
Downloading and installing remote packages.
Environment for https://raw.githubusercontent.com/snakemake-workflows/cyrcular-calling/v2.1.0/workflow/envs/bcftools.yaml created (location: .snakemake/conda/e9f1db15c0d3c38f4c28473ae5cdc670_)
Creating conda environment https://github.com/snakemake/snakemake-wrappers/raw/v1.25.0/bio/minimap2/aligner/environment.yaml...
Downloading and installing remote packages.
Environment for https://github.com/snakemake/snakemake-wrappers/raw/v1.25.0/bio/minimap2/aligner/environment.yaml created (location: .snakemake/conda/760088e00c13c025db1dc43a805800c8_)
Creating conda environment https://github.com/snakemake/snakemake-wrappers/raw/v1.25.0/bio/reference/ensembl-annotation/environment.yaml...
Downloading and installing remote packages.
Environment for https://github.com/snakemake/snakemake-wrappers/raw/v1.25.0/bio/reference/ensembl-annotation/environment.yaml created (location: .snakemake/conda/0fc8bd2d8acee9db91dcc38a4fd1bf12_)
Creating conda environment https://github.com/snakemake/snakemake-wrappers/raw/v1.25.0/bio/minimap2/index/environment.yaml...
Downloading and installing remote packages.
Environment for https://github.com/snakemake/snakemake-wrappers/raw/v1.25.0/bio/minimap2/index/environment.yaml created (location: .snakemake/conda/3609edbf7386a95c393e536852539e9f_)
Creating conda environment https://github.com/snakemake/snakemake-wrappers/raw/v1.25.0/bio/samtools/faidx/environment.yaml...
Downloading and installing remote packages.
Environment for https://github.com/snakemake/snakemake-wrappers/raw/v1.25.0/bio/samtools/faidx/environment.yaml created (location: .snakemake/conda/86ba35beb3d191069d6472b09bda5d32_)
Creating conda environment https://raw.githubusercontent.com/snakemake-workflows/cyrcular-calling/v2.1.0/workflow/envs/cyrcular.yaml...
Downloading and installing remote packages.
Environment for https://raw.githubusercontent.com/snakemake-workflows/cyrcular-calling/v2.1.0/workflow/envs/cyrcular.yaml created (location: .snakemake/conda/28015d0ac6596dc33c1e8186e4f0b356_)
Creating conda environment https://raw.githubusercontent.com/snakemake-workflows/cyrcular-calling/v2.1.0/workflow/envs/datavzrd.yaml...
Downloading and installing remote packages.
Environment for https://raw.githubusercontent.com/snakemake-workflows/cyrcular-calling/v2.1.0/workflow/envs/datavzrd.yaml created (location: .snakemake/conda/ba21bc8117aee787fe0a59947a7e5953_)
Creating conda environment https://raw.githubusercontent.com/snakemake-workflows/cyrcular-calling/v2.1.0/workflow/envs/pandas.yaml...
Downloading and installing remote packages.
Environment for https://raw.githubusercontent.com/snakemake-workflows/cyrcular-calling/v2.1.0/workflow/envs/pandas.yaml created (location: .snakemake/conda/4b444ac939a51643b8ca3c1a7874e512_)
Creating conda environment https://raw.githubusercontent.com/snakemake-workflows/cyrcular-calling/v2.1.0/workflow/envs/pigz.yaml...
Downloading and installing remote packages.
Environment for https://raw.githubusercontent.com/snakemake-workflows/cyrcular-calling/v2.1.0/workflow/envs/pigz.yaml created (location: .snakemake/conda/853ad1c780ed78c680a7892915d39095_)
Creating conda environment https://raw.githubusercontent.com/snakemake-workflows/cyrcular-calling/v2.1.0/workflow/envs/rbt.yaml...
Downloading and installing remote packages.
Environment for https://raw.githubusercontent.com/snakemake-workflows/cyrcular-calling/v2.1.0/workflow/envs/rbt.yaml created (location: .snakemake/conda/28dc58deed4445cc74232de79a311c81_)
Creating conda environment https://github.com/snakemake/snakemake-wrappers/raw/v1.25.0/bio/samtools/index/environment.yaml...
Downloading and installing remote packages.
Environment for https://github.com/snakemake/snakemake-wrappers/raw/v1.25.0/bio/samtools/index/environment.yaml created (location: .snakemake/conda/ca4357ab5cb510b9fda99ce03b644e13_)
Creating conda environment https://raw.githubusercontent.com/snakemake-workflows/cyrcular-calling/v2.1.0/workflow/envs/wget.yaml...
Downloading and installing remote packages.
Environment for https://raw.githubusercontent.com/snakemake-workflows/cyrcular-calling/v2.1.0/workflow/envs/wget.yaml created (location: .snakemake/conda/76d042288def0f75ec9e4fc0e00390ce_)
Using shell: /usr/bin/bash
Provided cores: 40
Rules claiming more threads will be scaled down.
Job stats:
job count
---------------------------------- -------
all 1
bcf_index 12
bcftools_concat 1
bcftools_sort 12
circle_bnds 1
circle_coverage_plot 1
circle_graph_plots 1
copy_graph_plots_for_datavzrd 4
copy_qc_plots_for_datavzrd 4
cyrcular_annotate_graph 1
cyrcular_generate_tables 1
datavzrd_circle_calls 1
download_gene_annotation 1
download_regulatory_annotation 1
download_repeatmasker_annotation 1
filter_overview_table 1
filter_varlociraptor 1
genome_faidx 1
get_bcf_header 1
get_genome 1
merge_fastqs 1
minimap2_bam 1
minimap2_index 1
reheader_filtered_bcf 1
render_datavzrd_config 1
samtools_index 1
scatter_candidates 1
sort_bcf_header 1
sort_bnd_bcfs 1
varlociraptor_alignment_properties 1
varlociraptor_call 12
varlociraptor_preprocess 12
total 82
Select jobs to execute...
Execute 5 jobs...
[Mon Aug 19 16:08:19 2024]
localrule download_regulatory_annotation:
output: resources/regulatory_annotation.gff3.gz
log: logs/download_regulatory_annotation.log
jobid: 14
reason: Missing output files: resources/regulatory_annotation.gff3.gz
resources: tmpdir=/tmp
Activating conda environment: .snakemake/conda/76d042288def0f75ec9e4fc0e00390ce_
[Mon Aug 19 16:08:19 2024]
localrule get_genome:
output: resources/homo_sapiens.GRCh38.107.fasta
log: logs/get-genome.log
jobid: 5
reason: Missing output files: resources/homo_sapiens.GRCh38.107.fasta
resources: tmpdir=/tmp
[Mon Aug 19 16:08:19 2024]
localrule download_gene_annotation:
output: resources/gene_annotation.gff3.gz
log: logs/download_gene_annotation.log
jobid: 13
reason: Missing output files: resources/gene_annotation.gff3.gz
resources: tmpdir=/tmp
[Mon Aug 19 16:08:19 2024]
localrule download_repeatmasker_annotation:
output: resources/repeat_masker.fa.out.gz
log: logs/download_repeatmasker_annotation.log
jobid: 15
reason: Missing output files: resources/repeat_masker.fa.out.gz
resources: tmpdir=/tmp
Activating conda environment: .snakemake/conda/76d042288def0f75ec9e4fc0e00390ce_
[Mon Aug 19 16:08:19 2024]
localrule merge_fastqs:
input: /home/swcho/1_ecDNA/3_data_for_WGS/nanopore_targeted/S23-10097_targeted_long_read_ERBB2.trimmed_Q7.fastq.gz
output: results/calling/merged/S23-10097_rmdup_single.fastq.gz
log: logs/merge-fastqs/S23-10097_rmdup_single.log
jobid: 10
reason: Missing output files: results/calling/merged/S23-10097_rmdup_single.fastq.gz
wildcards: sample=S23-10097_rmdup, read=single
resources: tmpdir=/tmp
Activating conda environment: .snakemake/conda/853ad1c780ed78c680a7892915d39095_
Activating conda environment: .snakemake/conda/0fc8bd2d8acee9db91dcc38a4fd1bf12_
Activating conda environment: .snakemake/conda/0fc8bd2d8acee9db91dcc38a4fd1bf12_
[Mon Aug 19 16:09:05 2024]
Finished job 13.
1 of 82 steps (1%) done
[Mon Aug 19 16:09:20 2024]
Finished job 14.
2 of 82 steps (2%) done
[Mon Aug 19 16:09:38 2024]
Finished job 15.
3 of 82 steps (4%) done
[Mon Aug 19 16:14:29 2024]
Finished job 10.
4 of 82 steps (5%) done
[Mon Aug 19 16:21:23 2024]
Finished job 5.
5 of 82 steps (6%) done
Select jobs to execute...
Execute 2 jobs...
[Mon Aug 19 16:21:24 2024]
localrule genome_faidx:
input: resources/homo_sapiens.GRCh38.107.fasta
output: resources/homo_sapiens.GRCh38.107.fasta.fai
log: logs/genome-faidx.log
jobid: 12
reason: Missing output files: resources/homo_sapiens.GRCh38.107.fasta.fai; Input files updated by another job: resources/homo_sapiens.GRCh38.107.fasta
resources: tmpdir=/tmp
[Mon Aug 19 16:21:24 2024]
localrule minimap2_index:
input: resources/homo_sapiens.GRCh38.107.fasta
output: resources/homo_sapiens.GRCh38.107.mmi
log: logs/minimap2_index/genome.log
jobid: 9
reason: Missing output files: resources/homo_sapiens.GRCh38.107.mmi; Input files updated by another job: resources/homo_sapiens.GRCh38.107.fasta
threads: 3
resources: tmpdir=/tmp
Activating conda environment: .snakemake/conda/86ba35beb3d191069d6472b09bda5d32_
Activating conda environment: .snakemake/conda/3609edbf7386a95c393e536852539e9f_
Activating conda environment: .snakemake/conda/86ba35beb3d191069d6472b09bda5d32_
[Mon Aug 19 16:21:49 2024]
Finished job 12.
6 of 82 steps (7%) done
[Mon Aug 19 16:23:11 2024]
Finished job 9.
7 of 82 steps (9%) done
Select jobs to execute...
Execute 1 jobs...
[Mon Aug 19 16:23:11 2024]
localrule minimap2_bam:
input: resources/homo_sapiens.GRCh38.107.mmi, results/calling/merged/S23-10097_rmdup_single.fastq.gz
output: results/calling/mapping/S23-10097_rmdup.bam
log: logs/minimap2/S23-10097_rmdup.log
jobid: 8
benchmark: benchmarks/minimap2/S23-10097_rmdup.txt
reason: Missing output files: results/calling/mapping/S23-10097_rmdup.bam; Input files updated by another job: results/calling/merged/S23-10097_rmdup_single.fastq.gz, resources/homo_sapiens.GRCh38.107.mmi
wildcards: sample=S23-10097_rmdup
threads: 20
resources: tmpdir=/tmp
Activating conda environment: .snakemake/conda/760088e00c13c025db1dc43a805800c8_
Activating conda environment: .snakemake/conda/760088e00c13c025db1dc43a805800c8_
[Mon Aug 19 17:24:56 2024]
Finished job 8.
8 of 82 steps (10%) done
Select jobs to execute...
Execute 2 jobs...
[Mon Aug 19 17:24:56 2024]
localrule varlociraptor_alignment_properties:
input: resources/homo_sapiens.GRCh38.107.fasta, resources/homo_sapiens.GRCh38.107.fasta.fai, results/calling/mapping/S23-10097_rmdup.bam
output: results/calling/alignment-properties/S23-10097_rmdup.json
log: logs/varlociraptor/estimate-alignment-properties/S23-10097_rmdup.log
jobid: 24
reason: Missing output files: results/calling/alignment-properties/S23-10097_rmdup.json; Input files updated by another job: results/calling/mapping/S23-10097_rmdup.bam, resources/homo_sapiens.GRCh38.107.fasta, resources/homo_sapiens.GRCh38.107.fasta.fai
wildcards: sample=S23-10097_rmdup
resources: tmpdir=/tmp
Activating conda environment: .snakemake/conda/2eef35d24a59f83df9961f5f70ab7240_
[Mon Aug 19 17:24:56 2024]
localrule samtools_index:
input: results/calling/mapping/S23-10097_rmdup.bam
output: results/calling/mapping/S23-10097_rmdup.bam.bai
log: logs/samtools_index/S23-10097_rmdup.log
jobid: 11
benchmark: benchmarks/samtools_index/S23-10097_rmdup.txt
reason: Missing output files: results/calling/mapping/S23-10097_rmdup.bam.bai; Input files updated by another job: results/calling/mapping/S23-10097_rmdup.bam
wildcards: sample=S23-10097_rmdup
threads: 12
resources: tmpdir=/tmp
Activating conda environment: .snakemake/conda/ca4357ab5cb510b9fda99ce03b644e13_
[Mon Aug 19 17:25:44 2024]
Finished job 11.
9 of 82 steps (11%) done
Select jobs to execute...
Execute 1 jobs...
[Mon Aug 19 17:25:44 2024]
localrule circle_bnds:
input: results/calling/mapping/S23-10097_rmdup.bam, results/calling/mapping/S23-10097_rmdup.bam.bai, resources/homo_sapiens.GRCh38.107.fasta, resources/homo_sapiens.GRCh38.107.fasta.fai
output: results/calling/candidates/S23-10097_rmdup.bcf, results/calling/graphs/S23-10097_rmdup.graph, results/calling/graphs/S23-10097_rmdup
log: logs/cyrcular/S23-10097_rmdup.log
jobid: 7
benchmark: benchmarks/cyrcular/S23-10097_rmdup.txt
reason: Missing output files: results/calling/candidates/S23-10097_rmdup.bcf, results/calling/graphs/S23-10097_rmdup.graph, results/calling/graphs/S23-10097_rmdup; Input files updated by another job: results/calling/mapping/S23-10097_rmdup.bam, resources/homo_sapiens.GRCh38.107.fasta, results/calling/mapping/S23-10097_rmdup.bam.bai, resources/homo_sapiens.GRCh38.107.fasta.fai
wildcards: sample=S23-10097_rmdup
threads: 4
resources: tmpdir=/tmp
Activating conda environment: .snakemake/conda/28015d0ac6596dc33c1e8186e4f0b356_
[Mon Aug 19 17:25:46 2024]
Error in rule circle_bnds:
jobid: 7
input: results/calling/mapping/S23-10097_rmdup.bam, results/calling/mapping/S23-10097_rmdup.bam.bai, resources/homo_sapiens.GRCh38.107.fasta, resources/homo_sapiens.GRCh38.107.fasta.fai
output: results/calling/candidates/S23-10097_rmdup.bcf, results/calling/graphs/S23-10097_rmdup.graph, results/calling/graphs/S23-10097_rmdup
log: logs/cyrcular/S23-10097_rmdup.log (check log file(s) for error details)
conda-env: /lustre/export/home/swcho/1_ecDNA/3_data_for_WGS/nanopore_targeted/cyrcular-calling/.snakemake/conda/28015d0ac6596dc33c1e8186e4f0b356_
threads: 12
resources: tmpdir=/tmp
Activating conda environment: .snakemake/conda/ca4357ab5cb510b9fda99ce03b644e13_
[Mon Aug 19 17:25:44 2024]
Finished job 11.
9 of 82 steps (11%) done
Select jobs to execute...
Execute 1 jobs...
[Mon Aug 19 17:25:44 2024]
localrule circle_bnds:
input: results/calling/mapping/S23-10097_rmdup.bam, results/calling/mapping/S23-10097_rmdup.bam.bai, resources/homo_sapiens.GRCh38.107.fasta, resources/homo_sapiens.GRCh38.107.fasta.fai
output: results/calling/candidates/S23-10097_rmdup.bcf, results/calling/graphs/S23-10097_rmdup.graph, results/calling/graphs/S23-10097_rmdup
log: logs/cyrcular/S23-10097_rmdup.log
jobid: 7
benchmark: benchmarks/cyrcular/S23-10097_rmdup.txt
reason: Missing output files: results/calling/candidates/S23-10097_rmdup.bcf, results/calling/graphs/S23-10097_rmdup.graph, results/calling/graphs/S23-10097_rmdup; Input files updated by another job: results/calling/mapping/S23-10097_rmdup.bam, resources/homo_sapiens.GRCh38.107.fasta, results/calling/mapping/S23-10097_rmdup.bam.bai, resources/homo_sapiens.GRCh38.107.fasta.fai
wildcards: sample=S23-10097_rmdup
threads: 4
resources: tmpdir=/tmp
Activating conda environment: .snakemake/conda/28015d0ac6596dc33c1e8186e4f0b356_
[Mon Aug 19 17:25:46 2024]
Error in rule circle_bnds:
jobid: 7
input: results/calling/mapping/S23-10097_rmdup.bam, results/calling/mapping/S23-10097_rmdup.bam.bai, resources/homo_sapiens.GRCh38.107.fasta, resources/homo_sapiens.GRCh38.107.fasta.fai
output: results/calling/candidates/S23-10097_rmdup.bcf, results/calling/graphs/S23-10097_rmdup.graph, results/calling/graphs/S23-10097_rmdup
log: logs/cyrcular/S23-10097_rmdup.log (check log file(s) for error details)
conda-env: /lustre/export/home/swcho/1_ecDNA/3_data_for_WGS/nanopore_targeted/cyrcular-calling/.snakemake/conda/28015d0ac6596dc33c1e8186e4f0b356_
shell:
cyrcular graph breakends results/calling/mapping/S23-10097_rmdup.bam --reference resources/homo_sapiens.GRCh38.107.fasta --min-read-depth 2 --min-split-reads 5 --max-paths-per-component 15 --max-deletion-length 10000 -t 4 --output results/calling/candidates/S23-10097_rmdup.bcf --graph results/calling/graphs/S23-10097_rmdup.graph --dot results/calling/graphs/S23-10097_rmdup 2> logs/cyrcular/S23-10097_rmdup.log
(one of the commands exited with non-zero exit code; note that snakemake uses bash strict mode!)
[Mon Aug 19 17:40:26 2024]
Finished job 24.
10 of 82 steps (12%) done
Shutting down, this might take some time.
Exiting because a job execution failed. Look above for error message
Complete log: .snakemake/log/2024-08-19T155240.245712.snakemake.log
WorkflowError:
At least one job did not complete successfully.
/usr/bin/bash: cyrcular: command not found
Thank you.
Hello, I'm a graduated student studying bioinformatics in South Korea.
I'm trying to do detect extrachromosomal DNA (ecDNA) in nanopore long-read targeted sequencing data.
I followed the workflow you described here.
I have tried two times of that workflow, but had same error in my job.
This is my standard log above
and this is the error log which place in
logs/cyrcular/I think it seems like the
cyrcular-callingpackages isn't installing properly from your github.Please check my logs and it will be grateful if you answer my question asap.
Thank you.