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JoVE Journal
Genetics

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

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

Article DOI: 10.3791/62607-v 05:51 min June 28th, 2021
June 28th, 2021

Chapters

Summary

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.

Tags

Digital-Droplet PCR Indels Mutations Genetically Modified Anopheline Mosquito Populations NHEJ Analysis DdPCR Sanger Sequencing Genetic Variation Field Analysis Experimental Systems Thai Pham Yang's Lab DdPCR Sample Mix Forward And Reverse Primers HEX Or FAM Probes DNA Water Vortexing Cartridge Oil Gasket Droplet Generator 96-well Plate
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