Seqanswers Leaderboard Ad

Collapse

Announcement

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

  • Using diff of polyA and ribozero prepped RNA?

    Hi all,
    I'm wondering if anyone knows if there are advantages to having both polyA and ribozero prepped RNAseq data? For example, one could potentially use the diff to find non-coding RNAs?

    I'm preparing an experiment with yeast and am planning on duplicates for each timepoint. I thought it might be interesting to do one set of replicates with polyA-enrichment and one set of replicates with ribozero enrichment. Not only would I still get the statistical power of duplicates, I could also get a direct measurement on non-polyA, non-ribosomal RNA. Am I wrong?

    Thanks!

  • #2
    There will be many differences between these two procedures that may or may not have to do with the actual polyA status of the transcripts. polyA selection generates a strong 3' bias among transcripts, depending on the quality (RIN) of the input RNA. Reports at meetings recently have indicated that the correlation between polyA selected and ribo-depleted libraries from the same input total RNA can be as low as 0.9, depending on the method used for ribo-depletion. Therefore, this could confound your method.

    That said, I do think it could work, you just have to keep possible bias of the rRNA-depletion method in mind when conducting downstream analyses.

    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