Seqanswers Leaderboard Ad

Collapse

Announcement

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

  • detect CRISPR edit using HRM

    Hello,
    I am trying to do knock-out in cell cultures using CRISPR.
    The cells are being transfected with a plasmid containing the essentials for CRISPR and a GFP marker. The transfection reates are quite low: about 20%, and out of those probably not all have been edited successfuly with CRISPR. After that DNA is extracted. Before I do FACS sorting for GFP and sequencing for detection of mutations/insertions/deletions, I wanted to just get an answer YES/NO if the CRISPR worked (don't care exactly what happened in the gene), a bit like pooling. So I thought to do HRM analysis to see if there is a difference between transfected cells and non transfected cells. The question is if it is possible to detect in HRM mutations even though not all cells and therefore not all the DNA is actually different from the control (non transfected cells).
    Bottom line, is it possible to detect differences in HRM if only about 10% of the template is edited/different between the samples.
    Thank you

  • #2
    Can you change your DNA with Crispr?
    My favorite hobby that I usually do when I have free time are web development. For more informationit chatbot solution companies singapore please visit the website here https://networkz.com.sg/best-chatbot...ies-singapore/

    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
    9 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