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Screening Sperm for the Rapid Isolation of Germline Edits in Zebrafish

Article DOI: 10.3791/64686-v • 05:55 min • February 10th, 2023
February 10th, 2023 •
1, 2, 1,2
1Department of Molecular Biosciences, The University of Texas at Austin, 2Department of Nutritional Sciences, The University of Texas at Austin

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CRISPR-Cas technologies have revolutionized the field of genome editing. However, finding and isolating the desired germline edit remains a major bottleneck. Therefore, this protocol describes a robust method for quickly screening F0 CRISPR-injected zebrafish sperm for germline edits using standard PCR, restriction digest, and gel electrophoresis techniques.

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Sperm Screening Germline Edits Zebrafish F0 CRISPR PCR Restriction Enzymes Gel Electrophoresis Indels Genome Edits In Vitro Fertilization Breeding Tanks Anesthetizing Fish Slotted Spoon Clean Paper Towels Sponge Anal Fins Cloaca Capillary Tube
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