Seqanswers Leaderboard Ad

Collapse

Announcement

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

  • Cluster density Illumina

    Hello, I would like to intoduce myself.
    We have been preparing and successfully sequencing a number of transcriptome libraries for PE Illumina sequencing.

    We obtained unexpectedly high cluster density with (supposedly) only 5nanoM of a transcriptome library loaded on a flow cell.
    The library was prepared using a NEB mRNA sample preparation kit and appeared normal when checked on a Bioanalyzer. <
    Has anyone else obtained (too) high cluster density with low sample input?

    Any comments welcome.

  • #2
    We switched to KAPA Q-PCR to quantify our libraries. This gives more reproducible cluster densitites.

    Comment


    • #3
      Predicting Cluster Densities

      Hi CatVincent,

      I recently went to an Illumina Sequencing seminar and from what I heard there, a majority of the users are using the Bioanalyzer in combination with another quantification method: PicoGreen (or RiboGreen in your case), NanoDrop, Qubit or qPCR to better predict cluster densities. It seemed universal that no one uses just the BioA beacuse it's not that predictive (I think someone mentioned within 25%).

      Our lab has been using PicoGreen in combination with BioAnalyzing and have been hitting our predicted cluster densities spot on! We're also going to start incorporating qPCR.

      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
      10 views
      0 likes
      Last Post seqadmin  
      Started by seqadmin, 03-22-2024, 10:03 AM
      0 responses
      51 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