Seqanswers Leaderboard Ad

Collapse

Announcement

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

  • ChIP-Seq library prep; trailing, primer dimer help?

    Hi,
    I've prepared ChIP-Seq libraries using NEBNext Multiplex Oligos for Illumina (Set 1, NEB #E7335) along with the NEBNext Ultra II DNA Library Prep Kit (NEB #E7645L). The samples will be sequenced on Novaseq 6000 S4, PE150, 30 million reads / sample.

    When I ran them on a bioanalyzer, they had trailing, primer dimer and adaptor dimer contamination. I did 1 PCR cycle followed by beads clean up and this helped in removing the adaptor dimers.
    I'm attaching a photo of the current bioanalyzer trace.

    Would it be ok to proceed with sequencing these libraries as they are?
    As far as I understand, the primer dimers will not be sequenced, since they don't contain any adapters. I'm not quite sure about the effect of the trailing that we see.
    If that's not ok, what are the consequences and what can I do?
    I was thinking of doing a size selection with the beads, but I'm not sure if that would result in a great loss of the libraries. Also, from what I read, the trailing could be carry over beads, if that's true and the libraries are placed on a magnet for a couple of minutes and the solution aspirated and collected, would that help?

    Thank you in advance.
    Attached Files
    Last edited by rozita; 11-25-2020, 05:45 AM. Reason: adding attachment

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