Unconfigured Ad

Collapse
X
 
  • Time
  • Show
Clear All
new posts
  • gwilymh
    Member
    • Dec 2011
    • 72

    #1

    Cufflinks returning elevated FPKM values for 'new' transcripts

    I am running RNA-Seq in a Tuxedo pipeline. In the final Cufflinks output, I am getting massively elevated FPKM values for some transcripts. These inflated values are only returned for the transcripts newly discovered by Cufflinks/TopHat (i.e. transcripts that were not previously annotated). The transcripts from annotated genes seemed normal.

    In one analysis, for example, the known genes had an average FPKM value of 214.5, and a maximum FPKM value of 147,473. The newly discovered transcripts, however, returned a mean FPKM of 139,234.6 and a maximum of 74,769,200; 2.5% of the new transcripts had vales greater than the maximum FPKM detected for the annotated genes. The results for the new transcripts clearly contain artifacts.

    My pipeline is below
    tophat \
    --min-anchor-length 10 \
    --splice-mismatches 1 \
    --min-intron-length 5 \
    --microexon-search \
    --fusion-search \
    --transcriptome-index=/gpfs/group1/f/flyinv/working_index/Dpse3_0_1_exons_ \
    -o "/gpfs/group1/f/flyinv/Outputs_TopHat/transcriptomeSequence_exons/AR_MSH126_Male" \
    /gpfs/group1/f/flyinv/working_index/Dpse3_0_1 \
    /gpfs/group1/f/flyinv/RNASeq/AR_MSH126_Male_1_sequence.txt \
    /gpfs/group1/f/flyinv/RNASeq/AR_MSH126_Male_2_sequence.txt \

    cufflinks \
    --output-dir "/gpfs/group1/f/flyinv/Outputs_CuffLinks/transcriptomeSequence_exons/AR_MSH126_Male" \
    --GTF-guide /gpfs/group1/f/flyinv/working_index/Dpse3_0_Exons.gff3 \
    --upper-quartile-norm \
    --min-intron-length 5 \
    --quiet \
    /gpfs/group1/f/flyinv/Outputs_TopHat/transcriptomeSequence_exons/AR_MSH126_Male/accepted_hits.bam \


    Does anyone know how or why this occurs? How can it be prevented? How can such artifacts be screened out of downstream analyses?

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...