Unconfigured Ad

Collapse
X
 
  • Filter
  • Time
  • Show
Clear All
new posts
  • Giles
    Member
    • Feb 2010
    • 39

    Multiplexing ChIP-seq

    I am aware of two Illumina kits that can do multiplexing: the "multiplexing sample preparation oligonucleotide kit" and the "TruSeq sample preparation kit". The steps between these procedures and the "ChIP-seq sample preparation kit" are qualitatively the same. However, it seems that the former two kits are designed for sequencing efforts that allow for large starting quantities of DNA (1-5ug). For ChIP-seq, you will be starting with more like 10ng.
    The question is, has anyone done multiplexing for ChIP-seq? If so, which kit did you use? And, did you tweak the final concentrations of any of the steps to prevent a massive excess of adapters during the ligation?
    Keith.
  • csquared
    Member
    • May 2008
    • 67

    #2
    Keith,

    We do it all the time with a custom kit that has some advantages to the TruSeq. Drop me a line and I can fill in the details. If you want to use the TruSeq kits, follow the protocol as written and just dilute the adaptors down 1:10 before ligation. All other steps can remain the same. The excess primer in the PCR isn't a problem.

    Shawn
    HudsonAlpha Institute for Biotechnology
    http://www.hudsonalpha.org/gsl

    Comment

    • Giles
      Member
      • Feb 2010
      • 39

      #3
      thanks for the info. That is exactly what I would have thought, but was hoping to get some confirmation.

      Comment

      • ETHANol
        Senior Member
        • Feb 2010
        • 308

        #4
        Any non-primary sequence heritable modification of genetic material. ChIP-SEQ, DNA methylation (Bisulfite-SEQ), chromatin modifications (methylation, acetylation, etc), non coding RNA.

        I've prepared several libraries using this protocol and it works great.

        csquared, I (and I bet others) would be interested in knowing the details. How about post it here.
        --------------
        Ethan

        Comment

        Latest Articles

        Collapse

        • SEQadmin2
          Nine Things a Sample Prep Scientist Thinks About Before Sequencing
          by SEQadmin2


          I’m not a sequencing expert. I’m a purification scientist who uses NGS to evaluate workflows my group develops. With this perspective, we think about the sample first and the NGS workflow second. The sequencer is an exceptionally honest reporter, but it can only report on what you give it, so whether you get clean, interpretable data from an NGS workflow is largely determined before you begin.


          Here are nine questions we think about, in roughly the order they matter, before...
          06-18-2026, 07:11 AM
        • SEQadmin2
          From Collection to Sequencing: Why Sample Preparation and Preservation Define Sequencing Data
          by SEQadmin2


          Data variability is still an issue in sequencing technologies despite the advances in reproducibility and accuracy of these platforms. But the problem does not originate in the sequencing itself, but in the previous steps, before the sample reaches the sequencer.


          The first step is collection, followed by preservation and sample preparation for analysis. Most scientists overlook those steps, but not being careful might just be skewing the experiment’s results.
          ...
          06-02-2026, 10:05 AM

        ad_right_rmr

        Collapse

        News

        Collapse

        Topics Statistics Last Post
        Started by SEQadmin2, 06-17-2026, 06:09 AM
        0 responses
        30 views
        0 reactions
        Last Post SEQadmin2  
        Started by SEQadmin2, 06-09-2026, 11:58 AM
        0 responses
        44 views
        0 reactions
        Last Post SEQadmin2  
        Started by SEQadmin2, 06-05-2026, 10:09 AM
        0 responses
        49 views
        0 reactions
        Last Post SEQadmin2  
        Started by SEQadmin2, 06-04-2026, 08:59 AM
        0 responses
        51 views
        0 reactions
        Last Post SEQadmin2  
        Working...