Seqanswers Leaderboard Ad

Collapse

Announcement

Collapse
No announcement yet.
X
 
  • Filter
  • Time
  • Show
Clear All
new posts

  • strange output from RSEM

    hi,
    I use RSEM (http://deweylab.biostat.wisc.edu/rsem/README.html) to analysis my RNA-seq data and want to summarize expression level of each RefSeq ID.
    the command I used is as below:
    rsem-calculate-expression -p 7 --fragment-length-min 51 --fragment-length-max 51 --fragment-length-mean 51 --fragment-length-sd 0
    BTW, the length of all my reads is 51bp

    I found that many isoform from the same gene got quite different TPM and FPKM. An example is as below:
    transcript_id gene_id length effective_length expected_count TPM FPKM IsoPct
    NM_001130147.1 CCDC77 2359 2309.00 317.27 16.83 11.21 100.00
    NM_001130148.1 CCDC77 2305 2255.00 2.23 0.12 0.08 100.00

    as you see. NM_001130147.1 and NM_001130148.1 are different isoform of CCDC77 and the mRNA seq is quite similar with each other. but the TPM as well as FPKM is quite different which I regard it as err.

    Does anyone meet the err? can you help me?

  • #2
    Why do you consider that to be an error? Typically there's only one major isoform.

    Comment


    • #3
      thank you for your replying, dpryan:

      because the mRNA seq of these two isoforms are nearly the same. Most of the reads mapped to NM_001130147.1 can also be mapped to NM_001130148.1. So I think the expression level of these two isoforms should be nearly the same.

      Am I right?

      Comment


      • #4
        No, just because most of the reads can map to both doesn't mean the expression was more or less the same. That's the entire point of programs like RSEM, have a read through the paper for details.

        Comment


        • #5
          Thank you, dpryan
          To be honest, I didn`t read the papper. Now I think it`s the time to read it.

          Comment

          Latest Articles

          Collapse

          • seqadmin
            Current Approaches to Protein Sequencing
            by seqadmin


            Proteins are often described as the workhorses of the cell, and identifying their sequences is key to understanding their role in biological processes and disease. Currently, the most common technique used to determine protein sequences is mass spectrometry. While still a valuable tool, mass spectrometry faces several limitations and requires a highly experienced scientist familiar with the equipment to operate it. Additionally, other proteomic methods, like affinity assays, are constrained...
            04-04-2024, 04:25 PM
          • seqadmin
            Strategies for Sequencing Challenging Samples
            by seqadmin


            Despite advancements in sequencing platforms and related sample preparation technologies, certain sample types continue to present significant challenges that can compromise sequencing results. Pedro Echave, Senior Manager of the Global Business Segment at Revvity, explained that the success of a sequencing experiment ultimately depends on the amount and integrity of the nucleic acid template (RNA or DNA) obtained from a sample. “The better the quality of the nucleic acid isolated...
            03-22-2024, 06:39 AM

          ad_right_rmr

          Collapse

          News

          Collapse

          Topics Statistics Last Post
          Started by seqadmin, 04-11-2024, 12:08 PM
          0 responses
          18 views
          0 likes
          Last Post seqadmin  
          Started by seqadmin, 04-10-2024, 10:19 PM
          0 responses
          22 views
          0 likes
          Last Post seqadmin  
          Started by seqadmin, 04-10-2024, 09:21 AM
          0 responses
          17 views
          0 likes
          Last Post seqadmin  
          Started by seqadmin, 04-04-2024, 09:00 AM
          0 responses
          49 views
          0 likes
          Last Post seqadmin  
          Working...
          X