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一种从任意 DNA 源进行大规模 gRNA 库生产的通用协议
JoVE Journal
Genetica
This content is Free Access.
JoVE Journal Genetica
A Universal Protocol for Large-scale gRNA Library Production from any DNA Source
DOI:

10:32 min

December 06, 2017

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Capitoli

  • 00:05Titolo
  • 00:48Digestion of DNA with MNase and Separation of DNA Fragments by PAGE
  • 03:05Isolation of DNA Fragments from PAGE Gels and Linker Generation
  • 04:26End Repair of Gel-purified Fragments and Linker Ligation
  • 05:32Size Selection and Assembly of Vector-insert Construct
  • 06:18Electroporation of TG1 Electrocompetent E. coli Cells
  • 08:38Results: a gRNA Library can be Generated from and Target DNA
  • 09:56Conclusion

Summary

Traduzione automatica

生成大型 gRNA 库的方法应简单、高效且具有成本效益。我们描述了一种基于目标 DNA 酶消化的 gRNA 库的生成协议。这种方法, CORALINA (全面 gRNA 图书馆一代通过控制核酸活动) 提出了一个替代昂贵的定制寡核苷酸合成。

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