Seqanswers Leaderboard Ad

Collapse

Announcement

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

  • a problem with RNA-Seq

    Hi All.

    I have problems with RNA-Seq. We used TruSeq kit to prepare libraries and Tophat to align reads. Take the following two genes as examples to show our problems.

    Click image for larger version

Name:	2012-9-19 11-37-32.png
Views:	1
Size:	21.2 KB
ID:	307914
    Almost no reads were mapped to exon 1 for Ptf1a.
    Click image for larger version

Name:	2012-9-19 11-45-24.png
Views:	1
Size:	26.2 KB
ID:	307915
    Many reads were mapped to introns for Rbpjl.

    At beginning, we thought it maybe the problem of alignment software. We tried GSNAP. However, the alignements were similar to Tophat.
    We suspected there might be some random errors in the experiments. However, when checking our another batch of RNA-Seq data which were done two months ago, we found the alignments were replicable between two batches.

    Does anyone have any ideas? Is there anything wrong with our experiments?

    Wish your help! Thanks very much! I appreciate it.

  • #2
    What was the RIN score of your RNA prior to TruSeq RNA prep?

    The lower the RIN score, the more 3' bias you would expect.
    --
    Phillip

    Comment

    Latest Articles

    Collapse

    • seqadmin
      Essential Discoveries and Tools in Epitranscriptomics
      by seqadmin




      The field of epigenetics has traditionally concentrated more on DNA and how changes like methylation and phosphorylation of histones impact gene expression and regulation. However, our increased understanding of RNA modifications and their importance in cellular processes has led to a rise in epitranscriptomics research. “Epitranscriptomics brings together the concepts of epigenetics and gene expression,” explained Adrien Leger, PhD, Principal Research Scientist...
      04-22-2024, 07:01 AM
    • 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

    ad_right_rmr

    Collapse

    News

    Collapse

    Topics Statistics Last Post
    Started by seqadmin, Today, 08:47 AM
    0 responses
    12 views
    0 likes
    Last Post seqadmin  
    Started by seqadmin, 04-11-2024, 12:08 PM
    0 responses
    60 views
    0 likes
    Last Post seqadmin  
    Started by seqadmin, 04-10-2024, 10:19 PM
    0 responses
    59 views
    0 likes
    Last Post seqadmin  
    Started by seqadmin, 04-10-2024, 09:21 AM
    0 responses
    54 views
    0 likes
    Last Post seqadmin  
    Working...
    X