zolotarovgl / GeneExt

GeneExt - Gene extension for improved scRNA-seq data counting
GNU General Public License v3.0
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problems in accepting annotation file #10

Open yurokohiya opened 4 months ago

yurokohiya commented 4 months ago

Hi,

I think I adjust the annotation file as required: image

and I ran like this: python geneext.py -g A.gtf -b all.bam" -o geneext.gtf

However, it failed. I wondered why: image

zolotarovgl commented 4 months ago

Dear, yurokohiya. The gene novel_gene2317... is missing the exons as indicated in the error message. You can try fixing your genome with agat tool before running GeneExt using the following command:

https://agat.readthedocs.io/en/latest/tools/agat_convert_sp_gxf2gxf.html

yurokohiya commented 4 months ago

Hi, this problem solved, but I also met another question in another case. I prepared the .gtf annotation file, and .bam file obtained from cellranger to run the pipeline, and geneExt seemed to run smoothly:

But there's nothing as the output. Then I noticed that I probably failed in creating index for .bam file, and it stuck here.

I wonder what caused this problem and how can I fix it?

At 2024-07-09 16:03:33, "Grygoriy Zolotarov, MD" @.***> wrote:

Dear, yurokohiya. The gene novel_gene2317... is missing the exons as indicated in the error message. You can try fixing your genome with agat tool before running GeneExt using the following command:

https://agat.readthedocs.io/en/latest/tools/agat_convert_sp_gxf2gxf.html

— Reply to this email directly, view it on GitHub, or unsubscribe. You are receiving this because you authored the thread.Message ID: @.***>

zolotarovgl commented 4 months ago

Hmm, why do you think it's a problem with .bam indexing? Have you tried indexing the .bam file yourself with samtools index ? Can you list the tmp folder contents? Usually, the source of most of such behaviour is a discrepancy between chromsomes in .bam file and gtf file - this causes bedtools commands to silently die and not easy to catch.