Unconfigured Ad

Collapse
X
 
  • Time
  • Show
Clear All
new posts
  • slowsmile
    Member
    • May 2011
    • 22

    #1

    can gff2 reference used in htseq-count?

    Dear all
    We are recently working with E.coli plasmid and tried to summarize the gene counts from our RNA-Seq samples.
    The short reads were mapped to E.coli plasmid using tophat which generated bam files accordingly.
    However, we were unable to obtain a gff3 version of our target plasmid genome, the best we can get was a gff2 file.
    Then I tried to use htseq-count to summarize the gene counts based on the gtf2 file, which I failed since the 9th column does not contain the required entry i.e. gene_id=XXXX, instead, they are gene_id "XXXX", which cannot be reconigized by the file.
    I then wrote a short script and manually changed the 9th field, by transforming gene_id "XXX" to gene_id="XXX".

    example of the transformed gff2 file is shown below
    gi|221630420|ref|NC_011917.1| RefSeq gene 1576 1788 . - . locus_tag=plLF82_002;db_xref=GeneID:7547171
    This time I sorted the reads and reran the htseq-count, however, I got another type of error:
    Error: The attribute string seems to contain mismatched quotes.
    [Exception type: ValueError, raised in __init__.py:168]
    Error occured in line 29 of file /media/max2/RNAseq-coloncancer/Microbiome/LF82_plasmid_new2.gff.
    Can anyway tell me what is going wrong with the procedure and whether there is a way to run htseq-count with gff2 reference file?

    Thanks a lot

Latest Articles

Collapse

  • SEQadmin2
    Multiomic Liquid Biopsy: What DNA/RNA Panels Reveal That DNA Alone Can’t
    by SEQadmin2



    Tumors contain complex mixtures of cell populations that evolve in response to treatment and host-specific factors. Tissue biopsies provide only static snapshots of a tumor’s state. But what if we could follow that evolution over time using a minimally invasive technique?

    Liquid biopsies can detect tumor-derived material circulating in the bloodstream, providing a way to study treatment response, detect emerging resistance, and discover new biomarkers. Circulating
    ...
    10-14-2026, 09:55 AM
  • SEQadmin2
    New Genomics Technologies Take Aim at Long-Standing Limits
    by SEQadmin2


    Researchers using sequencing and genomics tools often have to make trade-offs. They can choose between speed or scale, short reads or long-range information, or targeted panels or a view of the whole transcriptome. New technologies that have been released this year are built to address those tough choices.

    We asked six companies the same four questions to learn about their latest products. The new technologies bring a lot to the table, including rethinking sequencing
    ...
    09-28-2026, 10:25 AM

ad_right_rmr

Collapse

News

Collapse

Topics Statistics Last Post
Started by SEQadmin2, 10-22-2026, 11:13 AM
0 responses
26 views
0 reactions
Last Post SEQadmin2  
Started by SEQadmin2, 10-22-2026, 10:48 AM
0 responses
20 views
0 reactions
Last Post SEQadmin2  
Started by SEQadmin2, 10-15-2026, 10:52 AM
0 responses
36 views
0 reactions
Last Post SEQadmin2  
Started by SEQadmin2, 10-14-2026, 05:39 AM
0 responses
43 views
0 reactions
Last Post SEQadmin2  
Working...