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Targeted in Situ Mutagenesis of Histone Genes in Budding Yeast
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遗传学
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JoVE 杂志 遗传学
Targeted in Situ Mutagenesis of Histone Genes in Budding Yeast

Targeted in Situ Mutagenesis of Histone Genes in Budding Yeast

DOI:
10.3791/55263-v

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08:48 min

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January 26, 2017

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,
1Biology Department,Hendrix College

Chapters

  • 00:05Title
  • 00:46Generation and Purification of PCR Products Corresponding to Two Mutated and Partially Overlapping Histone Gene Fragments
  • 02:06Fusion PCR of the Two Partially Overlapping Fragments to Generate Full Size PCR Products for Integration
  • 03:20Co-transformation, Screen for 5-FOA-resistant Transformants, Purification of 5-FOA-resistant Colonies, and Loss of Backbone Plasmid
  • 05:11Results: In Situ Mutagenesis of HHT2 in Yeast
  • 07:09Conclusion

Summary

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A strategy for generating mutations in histone genes at their endogenous location in Saccharomyces cerevisiae is presented.

Tags

Targeted In Situ MutagenesisHistone GenesBudding YeastPCRFusion PCRTemplate DNAAgarose GelURA3 Gene5-FOA PlateReplica Plating

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