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. Author manuscript; available in PMC: 2020 May 1.
Published in final edited form as: Curr Protoc Mol Biol. 2017 Jul 5;119:31.9.1–31.9.22. doi: 10.1002/cpmb.42

Figure 7.

Figure 7

The HRMA results and 15% TBE gel used to analyze PCR products. (A) HRMA showing shifted melting curves with colored lines depicting injected embryos and the grey lines depicting the wild type uninjected control embryos. A greater deflection from the wt grey lines indicates higher mutation rates. (B) 15% TBE gel, which is used to analyze the PCR products and for visualizing DNA duplex formation in the CRISPR/Cas9 injected embryos vs wild type controls. The wt control lane shows a single PCR band while the Mutant embryo lanes show Heteroduplex formation induced by CRISPR injections. Heteroduplexes are indicated by the Red bracket. The Black asterisk * indicates the flh PCR product. The injected embryos were divided into 3 groups based on severity of phenotype penetrance. (C)The band intensity of the flh PCR product was analyzed using Image J software and plotted as a graph. The severity of the flh phenotype correlates directly to decreased levels of the flh PCR product.