How do I test guide RNA activity?

Given the time intensive nature of gene engineering, the relatively straightforward and quick (<1 week) process of gRNA validation can save weeks of cell culture and hours of bench time. Further to this, a clear idea of gRNA activity will provide insights into how many clones need to be screened to identify a positive.

There are now a variety of methodologies available to scientists looking to compare and validate guide RNA activity. Six of the most commonly used approaches are:

  1. Mismatch cleavage assay
  2. Sequence trace decomposition analysis
  3. Indel Detection by Amplicon Analysis (IDAA)
  4. Digital PCR
  5. Immunofluorescence analysis
  6. Clonal analysis

You'll find our complete guide to these approaches as well as tips for the Surveyor assay in our article here:

Six ways to validate guide RNA activity.

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