Unconfigured Ad

Collapse
X
 
  • Time
  • Show
Clear All
new posts
  • memento
    Member
    • Nov 2010
    • 49

    #1

    Solutions for multiplexing/barcoding

    Dear All,
    Can somebody point me out to a good and reasonably priced barcoding solution for illumina's MiSeq ? Are the adaptor sequences similar between MiSeq/HiSeq ? I found a barcoding generator by comai, but there is no protocol how to use it's output in a lab setting (probably very trivial question). I would assume that this is the cheapest solution. Bioo has a 96samp kit good for 768 reactions (question is how big rx they are talking about?) for 4995$. Are they the cheapest commercial solution ?
    Thanks in advance!
  • TonyBrooks
    Senior Member
    • Jun 2009
    • 303

    #2
    What application are you specifically looking at? RNA-Seq, DNA-Seq, ChIP-Seq, other...
    What level are you looking to plex (12, 48, 96, higher?)
    IMHO indexing primers are the most over-priced aspect of NGS. If you think you are going to be doing a lot of index library preparation, you may want to look at making your own.



    We ordered a set of indexes based on the above paper and use them regularly for both indexed (and non-indexed runs - you don't need to read the barcode). It works to be significantly cheaper per reaction than buying commercially, but there is obviously a largish initial outlay on the HPLC oligos. (Of course, you don't need to order all 96 in one go - one synthesis should provide enough for hundreds of reactions).
    Those oligo sequences work well on the GAIIx, HiSeq and MiSeq - just remember to use the custom index sequencing primer.

    There are also people on here dabbling with homebrew dual indexing, which is something we may look at in the future. It cuts the number of oligos you need to order down and will hopefully be scalable to >96 plex.
    Last edited by TonyBrooks; 10-15-2012, 04:03 AM.

    Comment

    • JackieBadger
      Senior Member
      • Mar 2009
      • 385

      #3
      for MiSeq amplicon work we use the 454 MIDs

      Comment

      • bhennuy
        Junior Member
        • May 2009
        • 6

        #4
        indexes sequence for multiplexed > 96

        Hello,
        We are interested in using dual indexing strategy for multiplexing more than 384 samples in one run on MiSeq instrument. Could we use the 48 indexes (6 bases) provided by Illumina for Truseq Small RNA sample prep as i7 indexes ? how to add 2 bases to these sequences to get unique 48 indexes ? We would use 8 i5 (as Nextera) indexes to reach 384 unique combinations. Do you think this strategy would work? Could we use more than 8 i5 indexes ?
        Thans for your help.

        Comment

        • TonyBrooks
          Senior Member
          • Jun 2009
          • 303

          #5
          You could look at this paper from Sanger

          They use 96-plex 8-mer barcodes. You could use those sequences for your i7 indexes.

          Comment

          • bhennuy
            Junior Member
            • May 2009
            • 6

            #6
            Thank you TonyBrooks for the info, I will read it carefully. Do you know why Illumina do not provide more i5 indexes to increase the number of multiplexes ?

            Comment

            Latest Articles

            Collapse

            • SEQadmin2
              Multiomic Liquid Biopsy: What DNA/RNA Panels Reveal That DNA Alone Can’t
              by SEQadmin2



              Tumors contain complex mixtures of cell populations that evolve in response to treatment and host-specific factors. Tissue biopsies provide only static snapshots of a tumor’s state. But what if we could follow that evolution over time using a minimally invasive technique?

              Liquid biopsies can detect tumor-derived material circulating in the bloodstream, providing a way to study treatment response, detect emerging resistance, and discover new biomarkers. Circulating
              ...
              10-14-2026, 09:55 AM
            • SEQadmin2
              New Genomics Technologies Take Aim at Long-Standing Limits
              by SEQadmin2


              Researchers using sequencing and genomics tools often have to make trade-offs. They can choose between speed or scale, short reads or long-range information, or targeted panels or a view of the whole transcriptome. New technologies that have been released this year are built to address those tough choices.

              We asked six companies the same four questions to learn about their latest products. The new technologies bring a lot to the table, including rethinking sequencing
              ...
              09-28-2026, 10:25 AM

            ad_right_rmr

            Collapse

            News

            Collapse

            Topics Statistics Last Post
            Started by SEQadmin2, 10-15-2026, 10:52 AM
            0 responses
            17 views
            0 reactions
            Last Post SEQadmin2  
            Started by SEQadmin2, 10-14-2026, 05:39 AM
            0 responses
            26 views
            0 reactions
            Last Post SEQadmin2  
            Started by SEQadmin2, 10-13-2026, 09:56 AM
            0 responses
            22 views
            0 reactions
            Last Post SEQadmin2  
            Started by SEQadmin2, 10-09-2026, 11:14 AM
            0 responses
            28 views
            0 reactions
            Last Post SEQadmin2  
            Working...