Seqanswers Leaderboard Ad

Collapse

Announcement

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

  • Illumina mRNAseq multiplexing

    Hi you all,

    we want to start mRNAseq with barcoded / multiplexed libraries on our Illumina GA II. Has anyone ever tried this? The standard Illumina multiplexing kit is designed for genomic DNA. Has anyone also used this for mRNAseq? Any modifications necessary?

    Or has anyone tried other approaches to this like using custom primers...

    Any help would be great,

    Fabian

  • #2
    Yes, you can definitely run indexed/multiplex libraries of RNA-Seq samples on the GA II (and HiSeq 2000). You just need to substitute multiplex adapters and PCR primers from the multiplexing kit for the adapters that are in the mRNA-Seq kit.

    Shawn Baker
    Market Manager, Expression and Epigenetics
    Illumina

    Comment


    • #3
      Agree with scbaker about hte multiplexing.

      We have used indexing by PCR primers for small RNA but I can't see why it wouldn't work for mRNA as you use the small RNA adapters with the strand specific protocol, although will need a bit of work.

      Comment


      • #4
        Works

        Hi,

        thanks to both of you and sorry for the late reply but we wantet to test your suggestions first.

        So the protocol works fine and we have successfully duplexed and triplexed samples using this approach.

        Best regards,

        Fabian

        Comment

        Latest Articles

        Collapse

        • 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
        • seqadmin
          Techniques and Challenges in Conservation Genomics
          by seqadmin



          The field of conservation genomics centers on applying genomics technologies in support of conservation efforts and the preservation of biodiversity. This article features interviews with two researchers who showcase their innovative work and highlight the current state and future of conservation genomics.

          Avian Conservation
          Matthew DeSaix, a recent doctoral graduate from Kristen Ruegg’s lab at The University of Colorado, shared that most of his research...
          03-08-2024, 10:41 AM

        ad_right_rmr

        Collapse

        News

        Collapse

        Topics Statistics Last Post
        Started by seqadmin, 03-27-2024, 06:37 PM
        0 responses
        13 views
        0 likes
        Last Post seqadmin  
        Started by seqadmin, 03-27-2024, 06:07 PM
        0 responses
        11 views
        0 likes
        Last Post seqadmin  
        Started by seqadmin, 03-22-2024, 10:03 AM
        0 responses
        53 views
        0 likes
        Last Post seqadmin  
        Started by seqadmin, 03-21-2024, 07:32 AM
        0 responses
        69 views
        0 likes
        Last Post seqadmin  
        Working...
        X