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Error with STAR-fusion FusionInspector #2

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kmavrommatis opened this issue Aug 16, 2018 · 7 comments
Open

Error with STAR-fusion FusionInspector #2

kmavrommatis opened this issue Aug 16, 2018 · 7 comments

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@kmavrommatis
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Hi,
I am running STAR fusion on a few 100s samples. All of them are 75bp PE reads.
Each sample runs on a cluster node with 8 cores, 64GB RAM without any other job running at the same time.
The total disk available to this job is over 1.5TB and by the end of the t
The command executed is

STAR-Fusion --left_fq $fastq1 --right_fq $fastq2 --genome_lib_dir /scratch/reference/genomes/Homo-sapiens/GRCh38.p2/STAR-Fusion --output_dir $outputDirectory/F1E000FC001.starfusion --CPU 8 --FusionInspector validate --tmpdir /scratch/tmp/kmavrommatis/STARfusion.human/

For few samples STAR-Fusion fails with the following error:

[91528000]   -retrieving read alignments for 118 spanning frags.
Error, cannot open file /scratch/tmp/kmavrommatis/STARfusion.human/31825/DA0000206/RNA-Seq/ProcessedData/STARfusion.human.hg38_1533331155/F1E000FC001.starfusion/FusionInspector-validate/fi_workdir/finspector.st
ar.cSorted.dupsMarked.bam at /opt/STAR-Fusion/FusionInspector/util/../PerlLib/SAM_reader.pm line 34.
        SAM_reader::_init('SAM_reader=HASH(0x24436b0)') called at /opt/STAR-Fusion/FusionInspector/util/../PerlLib/SAM_reader.pm line 24
        SAM_reader::new('SAM_reader', '/scratch/tmp/kmavrommatis/STARfusion.human/31825/DA0000206/RN...') called at /opt/STAR-Fusion/FusionInspector/util/get_fusion_SPANNING_reads_from_bam.from_chim_summary.pl
line 484
Traceback (most recent call last):
  File "/opt/STAR-Fusion/FusionInspector/FusionInspector", line 944, in <module>
    FusionInspector().run()
  File "/opt/STAR-Fusion/FusionInspector/FusionInspector", line 740, in run
    pipeliner.run()
  File "/opt/STAR-Fusion/FusionInspector/PyLib/Pipeliner.py", line 53, in run
    run_cmd(cmd.get_cmd())
  File "/opt/STAR-Fusion/FusionInspector/PyLib/Pipeliner.py", line 17, in run_cmd
    subprocess.check_call(cmd, shell=True)
  File "/usr/lib64/python2.7/subprocess.py", line 542, in check_call
    raise CalledProcessError(retcode, cmd)
subprocess.CalledProcessError: Command '/opt/STAR-Fusion/FusionInspector/util/get_fusion_SPANNING_reads_from_bam.from_chim_summary.pl --gtf_file /scratch/tmp/kmavrommatis/STARfusion.human/31825/DA0000206/RNA-Se
q/ProcessedData/STARfusion.human.hg38_1533331155/F1E000FC001.starfusion/FusionInspector-validate/fi_workdir/finspector.gtf --MIN_ALIGN_PER_ID 96 --bam /scratch/tmp/kmavrommatis/STARfusion.human/31825/DA0000206/
RNA-Seq/ProcessedData/STARfusion.human.hg38_1533331155/F1E000FC001.starfusion/FusionInspector-validate/fi_workdir/finspector.star.cSorted.dupsMarked.bam --junction_info /scratch/tmp/kmavrommatis/STARfusion.huma
n/31825/DA0000206/RNA-Seq/ProcessedData/STARfusion.human.hg38_1533331155/F1E000FC001.starfusion/FusionInspector-validate/fi_workdir/finspector.star.cSorted.dupsMarked.bam.fusion_junction_info > /scratch/tmp/kma
vrommatis/STARfusion.human/31825/DA0000206/RNA-Seq/ProcessedData/STARfusion.human.hg38_1533331155/F1E000FC001.starfusion/FusionInspector-validate/fi_workdir/finspector.star.cSorted.dupsMarked.bam.fusion_span_re
ads.sam' returned non-zero exit status 12
Error, cmd: /opt/STAR-Fusion/FusionInspector/FusionInspector --fusions star-fusion.fusion_predictions.abridged.tsv  --out_prefix finspector  --min_junction_reads 1  --min_novel_junction_support 3  --min_spannin
g_frags_only 5  --prep_for_IGV  --max_promiscuity 10  --out_dir /scratch/tmp/kmavrommatis/STARfusion.human/31825/DA0000206/RNA-Seq/ProcessedData/STARfusion.human.hg38_1533331155/F1E000FC001.starfusion/FusionIns
pector-validate  --genome_lib_dir /scratch/reference/genomes/Homo-sapiens/GRCh38.p2/STAR-Fusion  --CPU 8  --left_fq /scratch/tmp/kmavrommatis/STARfusion.human/31825/DA0000206/RNA-Seq/ProcessedData/CheckFastqQua
lityScore_1533331075/F1E000FC001_R1.fastq.gz --right_fq /scratch/tmp/kmavrommatis/STARfusion.human/31825/DA0000206/RNA-Seq/ProcessedData/CheckFastqQualityScore_1533331075/F1E000FC001_R2.fastq.gz  --annotate  di
ed with ret 256 No such file or directory at /opt/STAR-Fusion/PerlLib/Pipeliner.pm line 181.
        Pipeliner::run('Pipeliner=HASH(0x197b628)') called at /opt/STAR-Fusion/STAR-Fusion line 766

Any advise?

Thanks in advance for your help

@brianjohnhaas
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brianjohnhaas commented Aug 16, 2018 via email

@kmavrommatis
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kmavrommatis commented Aug 17, 2018

Hi,
thanks for the response.
The file/directory structure is not touched by any other operation.

The curious thing is that the "missing" dupsMarked.bam file is actually there, full length and with the correct permissions.
But after having verified that the file exists, I restart the job and it resumes from its last checkpoint and still failed at the same place with the same error

@brianjohnhaas
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brianjohnhaas commented Aug 17, 2018 via email

@stsergbg
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Hi!

It might be the wrong place since the problem was only STAR-Fusion related (not related to tutorial) but have you managed to identify the source of this particular problem? I've encountered the very same one with STAR-Fusion 1.8 - in an isolated docker container with no other processes PerlLib/SAM_reader.pm fails to read finspector.star.cSorted.dupsMarked.bam

Error, cannot open file /tmp/output_dir/FusionInspector-validate/fi_workdir/finspector.star.cSorted.dupsMarked.bam at /usr/local/src/STAR-Fusion/FusionInspector/util/../PerlLib/SAM_reader.pm line 34.

Sergey

@brianjohnhaas
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brianjohnhaas commented Oct 19, 2020 via email

@stsergbg
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Sorry for the late reply - this error seems to have been caused by insufficient storage space - increasing resources (disk quota and number of cores) helped me get rid of this error and execute everything successfully.

@brianjohnhaas
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brianjohnhaas commented Oct 25, 2020 via email

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