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Digital-Droplet PCR to Detect Indels Mutations in Genetically Modified Anopheline Mosquito Populations
Capitoli
- 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
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.
