Seqanswers Leaderboard Ad

Collapse

Announcement

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

  • TruSeq v3 Cluster Kits with TruSeq SBS kits?

    Hi all,

    Has anyone experimented with using TruSeq SBS kits (i.e. not v3 SBS kits) to sequence TruSeq v3 Cluster Kits and flow cells?. I have heard through the grapevine that it can be done and I was hoping that this might resolve a cluster kit / SBS kit mismatch problem.

    Thanks!

  • #2
    Didja try calling Illumina tech support?

    Comment


    • #3
      I haven't done it... but did ask the question. It can be done with no problem.
      The v3 cluster kit advancement is more reads (bigger flow cell/more clustering real estate), and improved GC resolution (chemistry changes). The v3 sequencing kit advancement is simply better stability, you can load 209 cycles of ICR, rather than split for a 2x101. TruSeq v1 seq kits can be used with v3 cluster kits with no reduction in q scores (presumably), you just have to split ICR for a 2x101

      Comment


      • #4
        Are there not issues with switching from v1 -> v3 requiring an initial "double priming" of fluids lines that blows an extra 4 cycles worth of reagents? I guess if you have not run v3 chemistry yet it doesn't matter. But if you were thinking about switching back and forth then this could be an issue.

        --
        Phillip

        Comment


        • #5
          Does this "double priming" have to happen once or every time you load a v1 vs. v3 flow cell? I actually buy 200 cycle kit (actually 202 cycle kits) and aliquot them into 4x 50 cycle kits (for which I can actually run 51 cycles—works like a charm). If double priming happens every time I switch then this will not be feasible...

          Comment


          • #6
            It is the SBS kit, not the flowcell that is the issue.

            From what we have been told, a double prime needs to be done every time one uses v3 chemistry in an instrument that used v1 chemistry for its previous run.

            The 200 cycle kits are supposed to be sufficient for a 4 (single prime)+ 101+ 7 +101 -- so roughly 213 cycles worth. A 50 cycle kit would (by the same logic) be 4 + 50 +7 = 61 cycles worth.

            --
            Phillip

            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
            7 views
            0 likes
            Last Post seqadmin  
            Started by seqadmin, Yesterday, 06:07 PM
            0 responses
            7 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
            66 views
            0 likes
            Last Post seqadmin  
            Working...
            X