Entering edit mode
4.8 years ago
dimitrischat
▴
210
when i try to run transcript_abundance.pl using RSEM
./align_and_estimate_abundance.pl --seqType fq --single '..x.fastq' --transcripts '.../Trinity.fasta' --output_dir '/..output' --est_method RSEM --aln_method bowtie2 --trinity_mode --prep_reference
it runs good up until :
samtools sort -n -@ 1 -m 1G -o bowtie2.bam.for_rsem.tmp.bam bowtie2.bam
[bam_sort_core] merging from 114 files and 1 in-memory blocks...
rsem-scan-for-paired-end-reads 1 bowtie2.bam.for_rsem.tmp.bam bowtie2.bam.for_rsem.bam
..................................................................................................
Finished!
Conversion is completed. bowtie2.bam.for_rsem.bam will be checked by 'rsem-sam-validator'.
rsem-sam-validator bowtie2.bam.for_rsem.bam
...................................................................................................................................................................................................................................................................................................................................................................................................................................................
The input file is valid!
then this error:
CMD: rsem-calculate-expression -p 4 --fragment-length-mean 200 --fragment-length-sd 80 --no-bam-output --bam bowtie2.bam.for_rsem.bam /Trinity_trimmed.fasta.RSEM RSEM
rsem-parse-alignments /trinity_trimmed/Trinity_trimmed.fasta.RSEM RSEM.temp/RSEM RSEM.stat/RSEM bowtie2.bam.for_rsem.bam 1 -tag XM
Cannot open /trinity_trimmed/Trinity_trimmed.fasta.RSEM.grp! It may not exist.
"rsem-parse-alignments /home/app/Trinity_trimmed.fasta.RSEM RSEM.temp/RSEM RSEM.stat/RSEM bowtie2.bam.for_rsem.bam 1 -tag XM" failed! Plase check if you provide correct parameters/options for the pipeline!
Error, cmd: rsem-calculate-expression -p 4 --fragment-length-mean 200 --fragment-length-sd 80 --no-bam-output --bam bowtie2.bam.for_rsem.bam /Trinity_trimmed.fasta.RSEM RSEM died with ret: 65280 at ./align_and_estimate_abundance.pl line 729.
I am having this same issue, 32 individuals into my dataset (not with all individuals). @dimitrischat how did you solve this?