Thank you Dear lieven.sterck. you solved my problem.
htseq-count returns 0 count for all
HI biostars
I'm struggling a problem. I want to make count with htseq-count,
10 bam files, aligned with hisat2 (alignment rate 60% - 80%),
sorted by samtools,
both fna file and gff downloaded from ncbi (one species),
head of bam file:
@HD VN:1.0 SO:coordinate
@SQ SN:chrNC_037328.1 LN:158534110
@SQ SN:chrNC_037329.1 LN:136231102
@SQ SN:chrNC_037330.1 LN:121005158
@SQ SN:chrNC_037331.1 LN:120000601
@SQ SN:chrNC_037332.1 LN:120089316
@SQ SN:chrNC_037333.1 LN:117806340
@SQ SN:chrNC_037334.1 LN:110682743
@SQ SN:chrNC_037335.1 LN:113319770
@SQ SN:chrNC_037336.1 LN:105454467
the first row of gff file is:
NC_037328.1 RefSeq region 1 158534110 . + . ID=NC_037328.1:1..158534110;Dbxref=taxon:9913;Name=1;breed=Hereford;chromosome=1;gbkey=Src;genome=chromosome;isolate=L1 Dominette 01449 registration number 42190680;mol_type=genomic DNA;sex=female;tissue-type=left lung
code I used for htseq:
htseq-count -t gene -i gene file.sorted.sam annotation.gff > file.txt
htseq-count -s yes -t gene -i ID file.sorted.sam annotation.gff > file.txt
even --idattr=gene and --idattr=exon and -i Parent -i transcript_id
but all of these just return 0 count or error.
would you please help me?
any answer is appreciated.
many thanks
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You have to make sure the sequence naming is exactly the same in your GFF as in you fasta file you use to align it to.
In you specific case : NC_037328.1 != chrNC_037328.1
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