Seqanswers Leaderboard Ad

Collapse

Announcement

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

  • Amplicon sequencing - fusion primers ordering

    Dear all,

    we would like to order fusion primers for 454 GS Junior amplicon sequencing.

    When consulting the manual, we are not sure if the primers need to have Phosphorothioated DNA bases on the ends and if B primer needs to be labeled with biotin (or is this true just for shotgun sequencing)?

    Thank you for your help.

  • #2
    Amplicon sequencing - fusion primers ordering

    Hi,

    The phosphorothioate bases are there to inhibit any exonuclease activity, so you can order them without. It did work in our hands but if you get any contamination you'll have a problem!

    The biotin on the B primer is necessary for the enrichment (enrichment beads are streptavidin coated magnetic beads) of the amplified beads, regardless the type of library.

    Comment


    • #3
      For the FLX, the primers do not need to be biotinylated; standard primers work fine. Biotin doesn't come into the process until the enrichment stage, and the biotin there comes from the enrichment primer, not from the primers you used to create your library. Furthermore, only one molecule of the original library you make with your primers will end up on the bead and even if it was biotinylated, the biotin molecule would be stripped off (along with that original molecule) when you add the melt solution.

      Comment

      Latest Articles

      Collapse

      • 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
      • seqadmin
        The Impact of AI in Genomic Medicine
        by seqadmin



        Artificial intelligence (AI) has evolved from a futuristic vision to a mainstream technology, highlighted by the introduction of tools like OpenAI's ChatGPT and Google's Gemini. In recent years, AI has become increasingly integrated into the field of genomics. This integration has enabled new scientific discoveries while simultaneously raising important ethical questions1. Interviews with two researchers at the center of this intersection provide insightful perspectives into...
        02-26-2024, 02:07 PM

      ad_right_rmr

      Collapse

      News

      Collapse

      Topics Statistics Last Post
      Started by seqadmin, 03-14-2024, 06:13 AM
      0 responses
      32 views
      0 likes
      Last Post seqadmin  
      Started by seqadmin, 03-08-2024, 08:03 AM
      0 responses
      71 views
      0 likes
      Last Post seqadmin  
      Started by seqadmin, 03-07-2024, 08:13 AM
      0 responses
      80 views
      0 likes
      Last Post seqadmin  
      Started by seqadmin, 03-06-2024, 09:51 AM
      0 responses
      68 views
      0 likes
      Last Post seqadmin  
      Working...
      X