Seqanswers Leaderboard Ad

Collapse

Announcement

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

  • Custom multiplex sequencing barcodes / indexes (8-mer)

    Hi all,

    I have synthesised a custom set of Illumina adaptors that use an 8-mer index to allow 384-plex multiplexing. I got these index sequences from the Nextera XT 384-plex index kit.

    I am looking to expand my index set beyond 384-plex. Preferably beyond 1000-plex. Does anyone have the sequences for an index set, with good balance, that would allow me to expand my custom adaptor multiplex size up to 1000 (or beyond)? I would prefer if the indexes were 8-mers but could try longer indexes if this allow expansion of the set...

    Thank you in advance for your help.

    Joe

  • #2
    Consider the 12 base index used in the following publication.

    The ongoing revolution in high-throughput sequencing continues to democratize the ability of small groups of investigators to map the microbial component of the biosphere. In particular, the coevolution of new sequencing platforms and new software tools allows data acquisition and analysis on an unp …

    Comment


    • #3
      There's a 16S amplicon paper (link below) with 32 i5 indices and 48 i7 indices which shoud be enough for 1536 libraries. The index sequences are in the supplementary material. The indices are dual 8bp similar to Nextera XT

      Comment


      • #4
        I suppose this is the two-step PCR approach? If so you could also add a short secondary index between the specific primer and the tail, to make it "double-dual" indexing. With two pairs of tailed, specific primers; one normal pair, and one pair with the secondary index inserted, you quadruple the number of possible combinations. In addition, the secondary index can also act as a heterogeneity spacer, thus helping with base diversity. Some manual demultiplexing will be required. We have done this a couple of times, it worked well.
        Jon

        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, Yesterday, 06:37 PM
        0 responses
        10 views
        0 likes
        Last Post seqadmin  
        Started by seqadmin, Yesterday, 06:07 PM
        0 responses
        9 views
        0 likes
        Last Post seqadmin  
        Started by seqadmin, 03-22-2024, 10:03 AM
        0 responses
        49 views
        0 likes
        Last Post seqadmin  
        Started by seqadmin, 03-21-2024, 07:32 AM
        0 responses
        67 views
        0 likes
        Last Post seqadmin  
        Working...
        X