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Created April 16, 2020 19:35
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$ nextflow run nf-core/rnaseq -profile docker --aligner hisat2 -r 1.4.2 --pseudo_aligner salmon --reads 'data/*_{1,2}_10k.fastq' --reverseStranded --max_cpus 6 --max_memory 32 -c config_bkv.txt
NOTE: Nextflow is not tested with Java 13.0.2 -- It's recommended the use of version 8 up to 12
N E X T F L O W ~ version 20.01.0
Launching `nf-core/rnaseq` [cheesy_northcutt] - revision: 3b6df9bd10 [1.4.2]
Extracting transcript fastas from genome fasta + gtf/gff
----------------------------------------------------
,--./,-.
___ __ __ __ ___ /,-._.--~'
|\ | |__ __ / ` / \ |__) |__ } {
| \| | \__, \__/ | \ |___ \`-._,-`-,
`._,._,'
nf-core/rnaseq v1.4.2
----------------------------------------------------
Pipeline Release : 1.4.2
Run Name : cheesy_northcutt
Reads : data/*_{1,2}_10k.fastq
Data Type : Paired-End
Strandedness : Reverse
Trimming : 5'R1: 0 / 5'R2: 0 / 3'R1: 0 / 3'R2: 0 / NextSeq Trim: 0
Aligner : HISAT2
Fasta Ref : refs/bkv.fa
Pseudo Aligner : Salmon
GTF Annotation : refs/GCF_000837865.1_ViralProj14074_genomic.gtf
Remove Ribosomal RNA: false
Biotype GTF field : gene_biotype
Save prefs : Ref Genome: No / Trimmed FastQ: No / Alignment intermediates: No
Max Resources : 32 memory, 6 cpus, 10d time per job
Container : docker - nfcore/rnaseq:1.4.2
Output dir : ./results
Launch dir : /Users/pgc92/projects/nextflow_nfcore_rnaseq/bkv
Working dir : /Users/pgc92/projects/nextflow_nfcore_rnaseq/bkv/work
Script dir : /Users/pgc92/.nextflow/assets/nf-core/rnaseq
User : pgc92
Config Profile : docker
----------------------------------------------------
executor > local (12)
[d1/21f419] process > get_software_versions [100%] 1 of 1 ✔
[98/e74888] process > makeBED12 [100%] 1 of 1 ✔
[50/08b28c] process > makeHisatSplicesites [100%] 1 of 1 ✔
[75/0b2dd9] process > makeHISATindex [100%] 1 of 1 ✔
[aa/e720f4] process > transcriptsToFasta [100%] 1 of 1 ✔
[f1/0491d4] process > makeSalmonIndex [100%] 1 of 1 ✔
[57/2acf16] process > fastqc [100%] 1 of 1 ✔
[97/26f5e8] process > trim_galore [100%] 1 of 1 ✔
[9e/24debe] process > hisat2Align [100%] 1 of 1 ✔
[2c/47d31b] process > hisat2_sortOutput [ 0%] 0 of 1
[- ] process > rseqc -
[- ] process > preseq -
[- ] process > markDuplicates -
[- ] process > qualimap -
[- ] process > dupradar -
[- ] process > featureCounts -
[- ] process > merge_featureCounts -
[- ] process > stringtieFPKM -
[- ] process > sample_correlation -
[03/39dd7c] process > salmon [ 0%] 0 of 1
[- ] process > salmon_tx2gene -
[- ] process > salmon_tximport -
[- ] process > salmon_merge -
[- ] process > multiqc -
[95/7e7e17] process > output_documentation [100%] 1 of 1 ✔
[HISAT2 index build] Available memory: 32 B
[HISAT2 index build] Less than 200 GB available, so NOT using splice sites and exons in HISAT2 index.
executor > local (12)
[d1/21f419] process > get_software_versions [100%] 1 of 1 ✔
[98/e74888] process > makeBED12 [100%] 1 of 1 ✔
[50/08b28c] process > makeHisatSplicesites [100%] 1 of 1 ✔
[75/0b2dd9] process > makeHISATindex [100%] 1 of 1 ✔
[aa/e720f4] process > transcriptsToFasta [100%] 1 of 1 ✔
[f1/0491d4] process > makeSalmonIndex [100%] 1 of 1 ✔
[57/2acf16] process > fastqc [100%] 1 of 1 ✔
[97/26f5e8] process > trim_galore [100%] 1 of 1 ✔
[9e/24debe] process > hisat2Align [100%] 1 of 1 ✔
[- ] process > hisat2_sortOutput -
[- ] process > rseqc -
[- ] process > preseq -
[- ] process > markDuplicates -
[- ] process > qualimap -
[- ] process > dupradar -
[- ] process > featureCounts -
[- ] process > merge_featureCounts -
[- ] process > stringtieFPKM -
[- ] process > sample_correlation -
[03/39dd7c] process > salmon [100%] 1 of 1, failed: 1 ✘
[- ] process > salmon_tx2gene -
[- ] process > salmon_tximport -
[- ] process > salmon_merge -
[- ] process > multiqc -
[95/7e7e17] process > output_documentation [100%] 1 of 1 ✔
[HISAT2 index build] Available memory: 32 B
[HISAT2 index build] Less than 200 GB available, so NOT using splice sites and exons in HISAT2 index.
[HISAT2 index build] Use --hisat_build_memory [small number] to skip this check.
[0;35m[nf-core/rnaseq] Pipeline completed with errors
executor > local (12)
[d1/21f419] process > get_software_versions [100%] 1 of 1 ✔
[98/e74888] process > makeBED12 [100%] 1 of 1 ✔
[50/08b28c] process > makeHisatSplicesites [100%] 1 of 1 ✔
[75/0b2dd9] process > makeHISATindex [100%] 1 of 1 ✔
[aa/e720f4] process > transcriptsToFasta [100%] 1 of 1 ✔
[f1/0491d4] process > makeSalmonIndex [100%] 1 of 1 ✔
[57/2acf16] process > fastqc [100%] 1 of 1 ✔
[97/26f5e8] process > trim_galore [100%] 1 of 1 ✔
[9e/24debe] process > hisat2Align [100%] 1 of 1 ✔
[- ] process > hisat2_sortOutput -
[- ] process > rseqc -
[- ] process > preseq -
[- ] process > markDuplicates -
[- ] process > qualimap -
[- ] process > dupradar -
[- ] process > featureCounts -
[- ] process > merge_featureCounts -
[- ] process > stringtieFPKM -
[- ] process > sample_correlation -
[03/39dd7c] process > salmon [100%] 1 of 1, failed: 1 ✘
[- ] process > salmon_tx2gene -
[- ] process > salmon_tximport -
[- ] process > salmon_merge -
[- ] process > multiqc -
[95/7e7e17] process > output_documentation [100%] 1 of 1 ✔
[HISAT2 index build] Available memory: 32 B
[HISAT2 index build] Less than 200 GB available, so NOT using splice sites and exons in HISAT2 index.
[HISAT2 index build] Use --hisat_build_memory [small number] to skip this check.
[0;35m[nf-core/rnaseq] Pipeline completed with errors
WARN: Killing pending tasks (1)
WARN: To render the execution DAG in the required format it is required to install Graphviz -- See http://www.graphviz.org for more info.
Error executing process > 'salmon (SRR11074361)'
Caused by:
Process `salmon (SRR11074361)` terminated with an error exit status (1)
Command executed:
salmon quant --validateMappings \
--seqBias --useVBOpt --gcBias \
--geneMap GCF_000837865.1_ViralProj14074_genomic.gtf \
--threads 6 \
--libType=ISR \
--index salmon_index \
-1 SRR11074361_1_10k_val_1.fq.gz -2 SRR11074361_2_10k_val_2.fq.gz \
-o SRR11074361
Command exit status:
1
Command output:
(empty)
Command error:
Version Info: ### PLEASE UPGRADE SALMON ###
### A newer version of salmon with bug fixes is available. ####
###
The newest version, available at https://github.com/COMBINE-lab/salmon/releases
contains new features, improvements, and bug fixes; please upgrade at your
earliest convenience.
###
Sign up for the salmon mailing list to hear about new versions, features and updates at:
https://oceangenomics.com/subscribe
###
### salmon (mapping-based) v0.14.1
### [ program ] => salmon
### [ command ] => quant
### [ validateMappings ] => { }
### [ seqBias ] => { }
### [ useVBOpt ] => { }
### [ gcBias ] => { }
### [ geneMap ] => { GCF_000837865.1_ViralProj14074_genomic.gtf }
### [ threads ] => { 6 }
### [ libType ] => { ISR }
### [ index ] => { salmon_index }
### [ mates1 ] => { SRR11074361_1_10k_val_1.fq.gz }
### [ mates2 ] => { SRR11074361_2_10k_val_2.fq.gz }
### [ output ] => { SRR11074361 }
Logs will be written to SRR11074361/logs
[2020-04-16 19:34:49.898] [jointLog] [info] Fragment incompatibility prior below threshold. Incompatible fragments will be ignored.
[2020-04-16 19:34:49.898] [jointLog] [info] Usage of --validateMappings implies use of minScoreFraction. Since not explicitly specified, it is being set to 0.65
[2020-04-16 19:34:49.898] [jointLog] [info] Usage of --validateMappings, without --hardFilter implies use of range factorization. rangeFactorizationBins is being set to 4
[2020-04-16 19:34:49.898] [jointLog] [info] Usage of --validateMappings implies a default consensus slack of 0.2. Setting consensusSlack to 0.2.
[2020-04-16 19:34:49.898] [jointLog] [info] parsing read library format
[2020-04-16 19:34:49.898] [jointLog] [info] There is 1 library.
[2020-04-16 19:34:49.965] [stderrLog] [info] Loading Suffix Array
[2020-04-16 19:34:49.960] [jointLog] [info] Loading Quasi index
[2020-04-16 19:34:49.963] [jointLog] [info] Loading 32-bit quasi index
[2020-04-16 19:34:49.970] [jointLog] [info] done
[2020-04-16 19:34:49.970] [jointLog] [info] Index contained 2 targets
[2020-04-16 19:34:49.968] [stderrLog] [info] Loading Transcript Info
[2020-04-16 19:34:49.970] [stderrLog] [info] Loading Rank-Select Bit Array
[2020-04-16 19:34:49.970] [stderrLog] [info] There were 2 set bits in the bit array
[2020-04-16 19:34:49.970] [stderrLog] [info] Computing transcript lengths
[2020-04-16 19:34:49.970] [stderrLog] [info] Waiting to finish loading hash
[2020-04-16 19:34:49.970] [stderrLog] [info] Done loading index
[2020-04-16 19:34:50.075] [jointLog] [warning] salmon was only able to assign 5 fragments to transcripts in the index, but the minimum number of required assigned fragments (--minAssignedFrags) was 10. This could be indicative of a mismatch between the reference and sample, or a very bad sample. You can change the --minAssignedFrags parameter to force salmon to quantify with fewer assigned fragments (must have at least 1).
Work dir:
/Users/pgc92/projects/nextflow_nfcore_rnaseq/bkv/work/03/39dd7c9568dfcf40e23ad037295928
Tip: you can replicate the issue by changing to the process work dir and entering the command `bash .command.run`
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