10XGenomics / cellranger

10x Genomics Single Cell Analysis
https://www.10xgenomics.com/support/software/cell-ranger
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mkref probelm #184

Closed AnnaAntonatouPap closed 8 months ago

AnnaAntonatouPap commented 1 year ago

Hello,

I am trying to build a custom mouse genome! I use the reference genome and gtf file from 10x and modified as described here: https://support.10xgenomics.com/single-cell-gene-expression/software/pipelines/latest/using/tutorial_mr#panel

My error message is the following

Creating new reference folder at /data/local/rajewsky/home/aantona/custome_genome_mouse_bgh/refdata-gex-mm10-2020-A_bigT_bGH/mm10-2020-A_BigT_bGH
...done

Writing genome FASTA file into reference folder...
...done

Indexing genome FASTA file...
Traceback (most recent call last):
  File "/data/local/rajewsky/home/aantona/yard/apps/cellranger-7.0.0/bin/rna/mkref", line 137, in <module>
    main()
  File "/data/local/rajewsky/home/aantona/yard/apps/cellranger-7.0.0/bin/rna/mkref", line 128, in main
    reference_builder.build_gex_reference()
  File "/data/local/rajewsky/home/aantona/yard/apps/cellranger-7.0.0/lib/python/cellranger/reference_builder.py", line 209, in build_gex_reference
    self.process_fasta()
  File "/data/local/rajewsky/home/aantona/yard/apps/cellranger-7.0.0/lib/python/cellranger/reference_builder.py", line 93, in process_fasta
    stdout = tk_subproc.check_output(
  File "/data/local/rajewsky/home/aantona/yard/apps/cellranger-7.0.0/lib/python/tenkit/log_subprocess.py", line 67, in check_output
    return subprocess.check_output(args, *wargs, **kwargs)
  File "/data/local/rajewsky/home/aantona/yard/apps/cellranger-7.0.0/external/anaconda/lib/python3.8/subprocess.py", line 415, in check_output
    return run(*popenargs, stdout=PIPE, timeout=timeout, check=True,
  File "/data/local/rajewsky/home/aantona/yard/apps/cellranger-7.0.0/external/anaconda/lib/python3.8/subprocess.py", line 516, in run
    raise CalledProcessError(retcode, process.args,
subprocess.CalledProcessError: Command '['/data/local/rajewsky/home/aantona/yard/apps/cellranger-7.0.0/external/anaconda/bin/samtools', 'faidx', '/data/local/rajewsky/home/aantona/custome_genome_mouse_bgh/refdata-gex-mm10-2020-A_bigT_bGH/mm10-2020-A_BigT_bGH/fasta/genome.fa']' returned non-zero exit status 1.
My command is 
`cellranger mkref --genome=mm10-2020-A_BigT_bGH  --fasta=mm10-2020-A_bigT_bGH.fa --genes=mm10-2020-A_bigT_bGH.gtf  --nthreads=64 --memgb=256 .`

and my folderlooks like this after running the cellranger mkref ( it stops when is creating the  genome.fa)
.
├── BigT_bgh.fa
├── bigT_bgh.gtf
├── cellRanger_mkfasta.sh
├── genes.gtf
├── genome.fa
**├── mm10-2020-A_BigT_bGH
│   └── fasta
│       └── genome.fa**
├── mm10-2020-A_bigT_bGH.fa
└── mm10-2020-A_bigT_bGH.gtf

Anna