Using a Fluorescent PCR-capillary Gel Electrophoresis Technique to Genotype CRISPR/Cas9-mediated Knockout Mutants in a High-throughput Format

The overall goal of this technique is to genotype CRISPR/Cas9 induced insertion deletion mutant clones in a high-throughput manner. This method can help answer key questions in the genetics field, such as the phenotypic effect of knocking out a particular gene in a SAIL model of choice. The main advantage of this technique is there is a minimal to high throughput screening that is multiflexible.

Though this method can provide insight into human cancer genetics, it can also be applied to the other systems, such as human, stem, and the primary cell, or the cells of non human origin. We first had the idea for this method when we performed short tandem repeats or STR typing assays. To begin the procedure, transfect the chosen cells with a plasmid, co expressing a fluorescent label cas9 and the single guide RNA.