Seqanswers Leaderboard Ad

Collapse

Announcement

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

  • CLIP/RIP-seq peak finding

    A question to all users that run CLIP/RIP-seq and then try to find peaks in their data.
    Since I couldn't find any specific tool for finding peaks in RNA-based data, I have decided to use the classical ChIP-seq tools (the logic is the same I think).
    I align reads to the genome (I don't really care about junction reads), and then run one of the ChIP-seq tools such as MACS.
    I would like to hear about the parameters people use when they run ChIP-seq tools on CLIP/RIP-seq data. For example many of the ChIP-seq tools use the "genome size" for random expectation calculations, and different genome sizes give different output.
    Do you use the transcriptome size or genome size when trying to find significant peaks?
    Any other issues that should be taken care of?
    Thanks
    Mali

  • #2
    I am by no means the expert on this topic but I have been reading about it a little here and there the last few days. From what I have read, using a ChIP-seq data analysis pipeline results in the identification of many false positives. I haven't seen any well documented workflows but you may want to check out this thread:
    Discussion of next-gen sequencing related bioinformatics: resources, algorithms, open source efforts, etc
    --------------
    Ethan

    Comment


    • #3
      Thanks ETHANol for your reply. Can you explain more why ChIP-seq pipeline is not suitable for RNA-based data, what is the reason for the high false-positive rate?
      I have RNA-IP sample and RNA-Input and would like to find enriched peaks. The idea is basically the same as with ChIP-seq.
      Mali

      Comment


      • #4
        I wish I could give you an answer but at this point I would probably just be polluting the forum with half-truths. Hopefully, someone with more experience on the topic will chime in.
        --------------
        Ethan

        Comment


        • #5
          We are interested in finding RNAs that interact with specific regions of chromatin. Do you know if there is a technique out there for this? Thanks.

          Comment


          • #6
            a) either you have a protein specifically associating with these loci, then use an IP followed by RNASeq

            b) otherwise the only regions in chromatin that can be purified so far (i know many people that tried others and failed) are telomers


            c) or do you work in yeast? then there might be other options.

            Comment

            Latest Articles

            Collapse

            • seqadmin
              Understanding Genetic Influence on Infectious Disease
              by seqadmin




              During the COVID-19 pandemic, scientists observed that while some individuals experienced severe illness when infected with SARS-CoV-2, others were barely affected. These disparities left researchers and clinicians wondering what causes the wide variations in response to viral infections and what role genetics plays.

              Jean-Laurent Casanova, M.D., Ph.D., Professor at Rockefeller University, is a leading expert in this crossover between genetics and infectious...
              09-09-2024, 10:59 AM
            • seqadmin
              Addressing Off-Target Effects in CRISPR Technologies
              by seqadmin






              The first FDA-approved CRISPR-based therapy marked the transition of therapeutic gene editing from a dream to reality1. CRISPR technologies have streamlined gene editing, and CRISPR screens have become an important approach for identifying genes involved in disease processes2. This technique introduces targeted mutations across numerous genes, enabling large-scale identification of gene functions, interactions, and pathways3. Identifying the full range...
              08-27-2024, 04:44 AM

            ad_right_rmr

            Collapse

            News

            Collapse

            Topics Statistics Last Post
            Started by seqadmin, 09-06-2024, 08:02 AM
            0 responses
            142 views
            0 likes
            Last Post seqadmin  
            Started by seqadmin, 09-03-2024, 08:30 AM
            0 responses
            146 views
            0 likes
            Last Post seqadmin  
            Started by seqadmin, 08-27-2024, 04:40 AM
            0 responses
            157 views
            0 likes
            Last Post seqadmin  
            Started by seqadmin, 08-22-2024, 05:00 AM
            0 responses
            399 views
            0 likes
            Last Post seqadmin  
            Working...
            X