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Detection of Homologous Recombination Intermediates via Proximity Ligation and Quantitative PCR in Saccharomyces cerevisiae
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Detection of Homologous Recombination Intermediates via Proximity Ligation and Quantitative PCR in Saccharomyces cerevisiae

Detection of Homologous Recombination Intermediates via Proximity Ligation and Quantitative PCR in Saccharomyces cerevisiae

DOI:
10.3791/64240-v

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07:55 min

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September 11, 2022

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, , ,
1Department of Microbiology & Molecular Genetics,University of California, Davis, 2Laboratory of Biology & Modeling of the Cell,École Normale Supérieure de Lyon, 3Department of Molecular & Cellular Biology,University of California, Davis

Chapters

  • 00:05Introduction
  • 00:50DLC Assay
  • 01:54Cell Spheroplasting, Lysis, and Restriction Site Restoration
  • 03:19Restriction Enzyme Digest and Intramolecular Ligation
  • 05:15Results: DLC and DLE Assay Analysis of D-Loops After DSB Induction
  • 07:02Conclusion

Summary

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The D-loop capture (DLC) and D-loop extension (DLE) assays utilize the principle of proximity ligation together with quantitative PCR to quantify D-loop formation, D-loop extension, and product formation at the site of an inducible double-stranded break in Saccharomyces cerevisiae.

Tags

Homologous RecombinationD-loopProximity LigationQuantitative PCRSaccharomyces CerevisiaeBRCA2Bloom SyndromeWerner SyndromeUV Cross-linkingSpheroplastingRestriction Enzyme Buffer

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