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

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

Article DOI: 10.3791/55586-v • 08:25 min • April 8th, 2017
April 8th, 2017 •
1, 1, 1, 1
1Cancer & Stem Cell Biology Programme, Duke-NUS Medical School

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The genotyping technique described here, which couples fluorescent polymerase chain reaction (PCR) to capillary gel electrophoresis, allows for high-throughput genotyping of nuclease-mediated knockout clones. It circumvents limitations faced by other genotyping techniques and is more cost effective than sequencing methods.

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Fluorescent PCR-capillary Gel Electrophoresis Genotype CRISPR/Cas9 Knockout Mutants High-throughput Format Genetics Field Phenotypic Effect SAIL Model Minimal To High Throughput Screening Multiflexible Human Cancer Genetics Stem Cells Primary Cells Non-human Cells Short Tandem Repeats (STR) Typing Assays Transfection Plasmid Fluorescent Label Cas9 Single Guide RNA Clones Penicillin Streptomycin Supplemented Growth Medium
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