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8.1 years ago
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Hello.
I used a ncRNA.fa to alignment using RNA-seq samples in TOPHAT
*FASTA:
>ENSSSCT00000024762.1 ncrna:novel chromosome:Sscrofa10.2:2:114672074:114672179:1 gene:ENSSSCG00000023111.1 gene_biotype:snRNA transcript_biotype:snRNA gene_symbol:U6 description:U6 spliceosomal RNA [Source:RFAM;Acc:RF00026]
GCGTCACTGTAGTAGCACACAGACTAAAACTGGAATGATACAGAGAAGATTAGCATAGAT
ACTGTGCCAGAATGGTGTATAAATTAAGGAAGCATTCTGTATTTTT*
My bam file its ok. There are sequence mapped. I look in igv the counts.
*HWI-7001432L:169:C7A9GANXX:2:1101:1151:88174 83 ENSSSCT00000019656.3 166 50 76M = 80 -162 GAAACAGCAGTGATAAAAATTAAGCCATGAACGAAAGTTTGACTAAGTTATATTAATTAGAGTTGGTAAATCTCGT 1GGGGGGGGEGGGGGGGGGGGGGGGGGDGGGGGGGGGGGGGGGGGGGGGGGGGGFF1GGGGGGGGGGGGGGBBB@A AS:i:0 XN:i:0 XM:i:0 XO:i:0 XG:i:0 NM:i:0 MD:Z:76 YT:Z:UU NH:i:1*
Im using a GTF from ensembl (that include no-coding genes)
* x havana exon 112829213 112829236 . + . exon_id "ENSSSCE00000248122"; exon_number "2"; exon_version "1"; gene_biotype "lincRNA"; gene_id "ENSSSCG00000030972"; gene_name "CH242-64G23.3"; gene_source "havana"; gene_version "1"; havana_transcript "OTTSUST00000012069"; havana_transcript_version "1"; transcript_biotype "lincRNA"; transcript_id "ENSSSCT00000033495"; transcript_name "CH242-64G23.3-001"; transcript_source "havana"; transcript_version "1"; tss_id "TSS25916";*
However, when I tried to calculted the counts by ht-seq... all transcripts appear 0 counts. I tried to change default parametres... but I did not succeed
Can anybody help me?
Thanks a lot!!