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JoVE Journal Genetics
Screening Sperm for the Rapid Isolation of Germline Edits in Zebrafish
 

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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résumé
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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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Keywords: Sperm Screening Germline Edits Zebrafish CRISPR F0 Indels Genome Edits In Vitro Fertilization PCR Gel Electrophoresis Anesthesia Cloaca Capillary Action Sodium Hydroxide Thermal Cycler Tris-HCl PCR Reaction Agarose Gel
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