Entering edit mode
18 months ago
demoraesdiogo2017
▴
110
Hello I am trying to analyze miRNA-seq data but I am having problems with the mapping. I always get pretty much 0 counts with the built in annotation file, so I got one from miRBase. However, I always get an error when I use it, it doesn't even run. I thought the error could have been in the structure of the file, so I downloaded one from GENCODEand compared both structures. They didn't seem different, but the one from GENCODE runs just like the built-in one (runs but gives zero counts). So I have no idea what is going on.
Here are the gff3 structures:
miRBase:
chr1 . miRNA_primary_transcript 12425986 12426106 . + . ID=MI0021869;Alias=MI0021869;Name=mmu-mir-6341
chr1 . miRNA 12426016 12426038 . + . ID=MIMAT0025084;Alias=MIMAT0025084;Name=mmu-miR-6341;Derives_from=MI0021869
chr1 . miRNA_primary_transcript 20679010 20679082 . + . ID=MI0000249;Alias=MI0000249;Name=mmu-mir-206
chr1 . miRNA 20679017 20679039 . + . ID=MIMAT0017004;Alias=MIMAT0017004;Name=mmu-miR-206-5p;Derives_from=MI0000249
GENCODE:
chr1 HAVANA gene 3073253 3074322 . + . ID=ENSMUSG00000102693.1;gene_id=ENSMUSG00000102693.1;gene_type=TEC;gene_status=KNOWN;gene_name=4933401J01Rik;level=2;havana_gene=OTTMUSG00000049935.1
chr1 HAVANA transcript 3073253 3074322 . + . ID=ENSMUST00000193812.1;Parent=ENSMUSG00000102693.1;gene_id=ENSMUSG00000102693.1;transcript_id=ENSMUST00000193812.1;gene_type=TEC;gene_status=KNOWN;gene_name=4933401J01Rik;transcript_type=TEC;transcript_status=KNOWN;transcript_name=4933401J01Rik-001;level=2;transcript_support_level=NA;tag=basic;havana_gene=OTTMUSG00000049935.1;havana_transcript=OTTMUST00000127109.1
chr1 HAVANA exon 3073253 3074322 . + . ID=exon:ENSMUST00000193812.1:1;Parent=ENSMUST00000193812.1;gene_id=ENSMUSG00000102693.1;transcript_id=ENSMUST00000193812.1;gene_type=TEC;gene_status=KNOWN;gene_name=4933401J01Rik;transcript_type=TEC;transcript_status=KNOWN;transcript_name=4933401J01Rik-001;exon_number=1;exon_id=ENSMUSE00001343744.1;level=2;transcript_support_level=NA;tag=basic;havana_gene=OTTMUSG00000049935.1;havana_transcript=OTTMUST00000127109.1
chr1 ENSEMBL gene 3102016 3102125 . + . ID=ENSMUSG00000064842.1;gene_id=ENSMUSG00000064842.1;gene_type=snRNA;gene_status=KNOWN;gene_name=Gm26206;level=3
chr1 ENSEMBL transcript 3102016 3102125 . + . ID=ENSMUST00000082908.1;Parent=ENSMUSG00000064842.1;gene_id=ENSMUSG00000064842.1;transcript_id=ENSMUST00000082908.1;gene_type=snRNA;gene_status=KNOWN;gene_name=Gm26206;transcript_type=snRNA;transcript_status=KNOWN;transcript_name=Gm26206-201;level=3;transcript_support_level=NA;tag=basic
any idea of what could be wrong?
What does your
featureCounts
command line look like?on the one that works. I change nothing when I use the one from mirBASE.