I had to deactivate the SortMeRNA step to make it work.
Would it be possible to add a failsafe for FASTQ integrity after each step that generates a FASTQ file? If necessary, fix the FASTQ file on the fly? I suggest this as a general solution.
Command used and terminal output
Command used:
nextflow run nf-core/rnaseq \
-r 3.17.0 \
-profile dkfz \
--input samples.csv \
--outdir ${PWD} \
--genome null \
--fasta ${REFERENCE_GENOME} \
--gtf ${REFERENCE_GTF} \
--additional_fasta ${REFERENCE_PHIX} \
--gencode \
--seq_center DKFZ \
--remove_ribo_rna \
--save_merged_fastq \
--save_reference \
--save_trimmed \
--save_align_intermeds \
--save_unaligned \
--save_non_ribo_reads \
--igenomes_ignore
Terminal output:
STAR version: 2.7.11b compiled: 2024-07-03T14:39:20+0000 :/opt/conda/conda-bld/star_1720017372352/work/source
Nov 20 22:22:10 ..... started STAR run
Nov 20 22:22:10 ..... loading genome
Nov 20 22:24:17 ..... processing annotations GTF
Nov 20 22:24:45 ..... inserting junctions into the genome indices
Nov 20 22:25:59 ..... started 1st pass mapping
Command error:
INFO: Environment variable SINGULARITYENV_TMPDIR is set, but APPTAINERENV_TMPDIR is preferred
INFO: Environment variable SINGULARITYENV_NXF_TASK_WORKDIR is set, but APPTAINERENV_NXF_TASK_WORKDIR is preferred
INFO: Environment variable SINGULARITYENV_NXF_DEBUG is set, but APPTAINERENV_NXF_DEBUG is preferred
EXITING because of FATAL ERROR in reads input: quality string length is not equal to sequence length
@ST-K00265:389:HMJW3BBXY:1:2223:9039:32244
ATAAAGTTGAAGGCTACAAGAAGACCAAGGAAGCTGTTTTGCTCCTTAAGAAACTTAAAGCCTGGAATGATATCAAAAAGGTCTATGCCTCTCAGCGAATG
<A
SOLUTION: fix your fastq file
Nov 20 22:31:36 ...... FATAL ERROR, exiting
Relevant files
No response
System information
Nextflow version: 24.10.1
Hardware: HPC
Executer: lsf
Container engine: Singularity
OS: CentOS 7
Version of nf-core/rnaseq: 3.17.0
Description of the bug
This issue was discussed in SortMeRNA repository already (https://github.com/sortmerna/sortmerna/issues/407).
STAR failed for one sample due to the sequence and quality lengths mismatching for a read. After TrimGalore I have this
which becomes this after SortMeRNA:
I had to deactivate the SortMeRNA step to make it work.
Would it be possible to add a failsafe for FASTQ integrity after each step that generates a FASTQ file? If necessary, fix the FASTQ file on the fly? I suggest this as a general solution.
Command used and terminal output
Relevant files
No response
System information
Nextflow version: 24.10.1 Hardware: HPC Executer: lsf Container engine: Singularity OS: CentOS 7 Version of nf-core/rnaseq: 3.17.0