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CRISPR Guide RNA Cloning for Mammalian Systems
JoVE Journal
Genética
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JoVE Journal Genética
CRISPR Guide RNA Cloning for Mammalian Systems

CRISPR Guide RNA Cloning for Mammalian Systems

DOI:
10.3791/57998-v

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06:48 min

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October 02, 2018

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, , , , , , ,
1Wyss Institute for Biologically Inspired Engineering,Harvard University, 2Department of Genetics,Harvard Medical School, 3Department of Pathology,Massachusetts General Hospital, 4Institute for Medical Engineering & Science,Massachusetts Institute of Technology, 5Synthetic Biology Center,Massachusetts Institute of Technology, 6Department of Biological Engineering,Massachusetts Institute of Technology, 7Broad Institute

Capítulos

  • 00:04Título
  • 00:25Preparation of the Primers and Expression Vectors
  • 02:05Ligating the Annealed gRNA Oligonucleotides
  • 02:59Transformation of E. coli
  • 04:03PCR Analysis
  • 05:35Results: CRISPR Guided RNA Cloning
  • 06:23Conclusion

Summary

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Here, a simple, efficient, and cost-effective method of sgRNA cloning is outlined.

Tags

CRISPRGuide RNACloningMammalian SystemPlasmidSingle-guide RNAOligonucleotideLigationE. ColiCompetent CellsTransformation

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