Header Leaderboard Ad

Collapse

Library with strong secondary structures

Collapse

Announcement

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

  • Library with strong secondary structures

    What is known about the strand displacement activity of the DNA polymerase Illumina uses for bridge amplification? I have a library with strong secondary structures and I am worried that clusters do not form well due to the secondary structures. Does anyone have any tips to improve cluster formation for such a library?

  • #2
    Anecdotally we find MiSeq to be the best sequencer for odd samples. You best bet is to sequence this library there. You can always a run a nano flowcell at a much reduced cost to test before you run full library.

    Comment


    • #3
      You can also try a 95C denaturation (instead of the usual NaOH denaturation). 1 minute at that temperature (in RSB or EB should be fine), just make sure you have a decent volume there so you don't evaporate everything. TruSeq Custom Amplicon product does this method to make the libraries nice and single stranded for cluster generation.

      Comment

      Latest Articles

      Collapse

      • seqadmin
        A Brief Overview and Common Challenges in Single-cell Sequencing Analysis
        by seqadmin


        ​​​​​​The introduction of single-cell sequencing has advanced the ability to study cell-to-cell heterogeneity. Its use has improved our understanding of somatic mutations1, cell lineages2, cellular diversity and regulation3, and development in multicellular organisms4. Single-cell sequencing encompasses hundreds of techniques with different approaches to studying the genomes, transcriptomes, epigenomes, and other omics of individual cells. The analysis of single-cell sequencing data i...

        01-24-2023, 01:19 PM
      • seqadmin
        Introduction to Single-Cell Sequencing
        by seqadmin
        Single-cell sequencing is a technique used to investigate the genome, transcriptome, epigenome, and other omics of individual cells using high-throughput sequencing. This technology has provided many scientific breakthroughs and continues to be applied across many fields, including microbiology, oncology, immunology, neurobiology, precision medicine, and stem cell research.

        The advancement of single-cell sequencing began in 2009 when Tang et al. investigated the single-cell transcriptomes
        ...
        01-09-2023, 03:10 PM

      ad_right_rmr

      Collapse
      Working...
      X