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Digital-Droplet PCR to Detect Indels Mutations in Genetically Modified Anopheline Mosquito Populations
JoVE Journal
Genetics
This content is Free Access.
JoVE Journal Genetics
Digital-Droplet PCR to Detect Indels Mutations in Genetically Modified Anopheline Mosquito Populations
DOI:

05:51 min

June 28, 2021

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Chapters

  • 00:04Introduction
  • 00:48PCR Preparation and Droplet Generation
  • 02:18PCR
  • 03:03Droplet Reading
  • 03:31Analysis
  • 04:49Results: Implications of ddPCR for NHEJ Identification and Quantification in Transgenic Anopheline Mosquitoes
  • 05:27Conclusion

Summary

Automatic Translation

This protocol provides the steps from DNA extraction to experimental set-up for digital droplet PCR (ddPCR), including analysis for the identification and quantification of non-homologous end-joining (NHEJ) events at target sites following gRNA-induced Cas9 cleavage and DNA repair. Other uses of this method include applications such as polymorphism detection and gene-editing variant verification.

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