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CRISPR/Cas9 使用dpy-10联合 CRISPR 筛选标记和组装的核糖核蛋白复合物编辑C. elegans rbm-3.2基因。
JoVE Journal
Genetica
This content is Free Access.
JoVE Journal Genetica
CRISPR/Cas9 Editing of the C. elegans rbm-3.2 Gene using the dpy-10 Co-CRISPR Screening Marker and Assembled Ribonucleoprotein Complexes.
DOI:

07:46 min

December 11, 2020

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Capitoli

  • 00:04Introduction
  • 00:53C. elegans Gonad Preparation and Microinjection
  • 01:56Injected Worm Recovery and Transfer
  • 02:37PickingC. elegans for Screening
  • 03:17Single Worm Lysis and PCR
  • 04:22Restriction Digestion and Agarose Gel Electrophoresis
  • 05:08Identification of Positively Edited and Homozygous Edited Worms
  • 06:01Results: Representative C. elegans rbm-3.2 Gene Editing and Screening
  • 07:10Conclusion

Summary

Traduzione automatica

此协议的目标是使用组装的核糖核蛋白复合物和dpy-10共同 CRISPR 标记进行无缝的 CRISPR/Cas9 编辑,以便进行筛选。 此协议可用于在C. elegans中进行各种基因修改,包括插入、删除、基因替换和 codon 替代。

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