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pliu55 avatar pliu55 commented on September 12, 2024

@sopenaml, what command did you use when you got this error?

from rsem.

sopenaml avatar sopenaml commented on September 12, 2024

It's a bit convoluted, but I'm running the nfcore/rnaseq pipeline using STAR-rsem to align/quantify counts per gene and my fastq files are quite big. Below is the error obtained that also includes the step where the error occurred.

Error executing process > 'NFCORE_RNASEQ:RNASEQ:QUANTIFY_RSEM:RSEM_CALCULATEEXPRESSION (HepG2_Rep3)'

Caused by:
 Process `NFCORE_RNASEQ:RNASEQ:QUANTIFY_RSEM:RSEM_CALCULATEEXPRESSION (HepG2_Rep3)` terminated with an error exit status (255)

Command executed:

 INDEX=`find -L ./ -name "*.grp" | sed 's/\.grp$//'`
 rsem-calculate-expression \
   --num-threads 12 \
   --temporary-folder ./tmp/ \
   --strandedness forward \
   --paired-end \
   --star --star-output-genome-bam --star-gzipped-read-file --estimate-rspd --seed 1 \
   HepG2_Rep3_1_val_1.fq.gz HepG2_Rep3_2_val_2.fq.gz \
   $INDEX \
   HepG2_Rep3
  
 cat <<-END_VERSIONS > versions.yml
 "NFCORE_RNASEQ:RNASEQ:QUANTIFY_RSEM:RSEM_CALCULATEEXPRESSION":
   rsem: $(rsem-calculate-expression --version | sed -e "s/Current version: RSEM v//g")
   star: $(STAR --version | sed -e "s/STAR_//g")
 END_VERSIONS

Command exit status:
 255

Command output:
 STAR --genomeDir ./rsem --outSAMunmapped Within --outFilterType BySJout --outSAMattributes NH HI AS NM MD --outFilterMultimapNmax 20 --outFilterMismatchNmax 999 --outFilterMismatchNoverLmax 0.04 --alignIntronMin 20 --alignIntronMax 1000000 --alignMatesGapMax 1000000 --alignSJoverhangMin 8 --alignSJDBoverhangMin 1 --sjdbScore 1 --runThreadN 12 --genomeLoad NoSharedMemory --outSAMtype BAM Unsorted --quantMode TranscriptomeSAM --outSAMheaderHD @HD VN:1.4 SO:unsorted --outFileNamePrefix ./tmp//HepG2_Rep3 --readFilesCommand zcat --readFilesIn HepG2_Rep3_1_val_1.fq.gz HepG2_Rep3_2_val_2.fq.gz
 	STAR --genomeDir ./rsem --outSAMunmapped Within --outFilterType BySJout --outSAMattributes NH HI AS NM MD --outFilterMultimapNmax 20 --outFilterMismatchNmax 999 --outFilterMismatchNoverLmax 0.04 --alignIntronMin 20 --alignIntronMax 1000000 --alignMatesGapMax 1000000 --alignSJoverhangMin 8 --alignSJDBoverhangMin 1 --sjdbScore 1 --runThreadN 12 --genomeLoad NoSharedMemory --outSAMtype BAM Unsorted --quantMode TranscriptomeSAM --outSAMheaderHD @HD VN:1.4 SO:unsorted --outFileNamePrefix ./tmp//HepG2_Rep3 --readFilesCommand zcat --readFilesIn HepG2_Rep3_1_val_1.fq.gz HepG2_Rep3_2_val_2.fq.gz
 	STAR version: 2.7.10a  compiled: 2022-01-14T18:50:00-05:00 :/home/dobin/data/STAR/STARcode/STAR.master/source
 Jul 12 17:07:07 ..... started STAR run
 Jul 12 17:07:07 ..... loading genome
 Jul 12 17:07:43 ..... started mapping
 "STAR --genomeDir ./rsem --outSAMunmapped Within --outFilterType BySJout --outSAMattributes NH HI AS NM MD --outFilterMultimapNmax 20 --outFilterMismatchNmax 999 --outFilterMismatchNoverLmax 0.04 --alignIntronMin 20 --alignIntronMax 1000000 --alignMatesGapMax 1000000 --alignSJoverhangMin 8 --alignSJDBoverhangMin 1 --sjdbScore 1 --runThreadN 12 --genomeLoad NoSharedMemory --outSAMtype BAM Unsorted --quantMode TranscriptomeSAM --outSAMheaderHD @HD VN:1.4 SO:unsorted --outFileNamePrefix ./tmp//HepG2_Rep3 --readFilesCommand zcat --readFilesIn HepG2_Rep3_1_val_1.fq.gz HepG2_Rep3_2_val_2.fq.gz" failed! Plase check if you provide correct parameters/options for the pipeline!

Command error:
  
 EXITING because of fatal error: buffer size for SJ output is too small
 Solution: increase input parameter --limitOutSJcollapsed
  
 Jul 12 17:35:08 ...... FATAL ERROR, exiting

from rsem.

pliu55 avatar pliu55 commented on September 12, 2024

STAR's default buffer size for SJ output is usually fine for human data. Just for testing and perhaps as a temporary fix, would you like to replace rsem-calculate-expression's line 489 by the following?

" --outSAMheaderHD \@HD VN:1.4 SO:unsorted --limitOutSJcollapsed 2000000 ".

assuming you want to increase it to 2000000.

from rsem.

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