Seqanswers Leaderboard Ad

Collapse

Announcement

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

  • Is Nextera useful for Pac bio in future?

    Hi There,

    I am working with Nextera workflow. I was wondering if Nextera library prep can be somehow extended to Pac bio platform. My understanding is Nextera insert size is much smaller than what Pac bio can sequence. Is there a way Nextera transposon can fragment a bigger size insert, add adapters and then it gets sequenced on Pac bio.

    This way, we can use advantage of both the technologies, low imput starting material and sequencing it real-time.

    Any suggestions?

  • #2
    I suspect that if you were to reduce the amount of transposase compared to DNA then you would see an increase in the bias of the fragmentation, as the preferred sites would use up the low amount of reagent.

    Nextera is nice because it allows you to skip end-repair and ligation of adapters. But if you want to make really long DNA fragments, it is probably preferable to just end repair and ligate and not add a fragmentation step.
    Providing nextRAD genotyping and PacBio sequencing services. http://snpsaurus.com

    Comment


    • #3
      It can be done in two ways. One is to do tagmentation with more input DNA or reduced enzyme to obtain large tagmented DNA for amplification as suggested by SNPsaurus. Other is to tagment DNA as before but fill in the gap and end repair without amplification. One also can introduce size selection either way. Nextera mate pair tagmentation system also can be used but it is less cost effective.

      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
      8 views
      0 likes
      Last Post seqadmin  
      Started by seqadmin, Yesterday, 06:07 PM
      0 responses
      8 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