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筛选精子以快速分离斑马鱼的种系编辑

JoVE Journal
Genética

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JoVE Journal Genética

Screening Sperm for the Rapid Isolation of Germline Edits in Zebrafish

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筛选精子以快速分离斑马鱼的种系编辑

DOI:

10.3791/64686-v

•

05:55 min

•

February 10, 2023

•

Brittney Voigt, Ryoko Minowa, Ryan S. Gray²

¹Department of Molecular Biosciences,The University of Texas at Austin, ²Department of Nutritional Sciences,The University of Texas at Austin

Capítulos

00:04Introduction
00:50Fish Sperm Collection
02:11DNA Extraction and Amplification
03:14PCR Amplicon Separation by Gel Electrophoresis
04:20Results: Mutation Analysis of Sperm
05:16Conclusion

Summary

Tadução automática

CRISPR-Cas技术彻底改变了基因组编辑领域。然而，寻找和分离所需的种系编辑仍然是一个主要瓶颈。因此，该协议描述了一种使用标准PCR，限制性酶切和凝胶电泳技术快速筛选F0 CRISPR注射斑马鱼精子以进行种系编辑的稳健方法。

Tags

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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