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通过邻近连接和定量PCR检测酿酒酵母同源重组中间体

JoVE Journal
Genética

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JoVE Journal Genética

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

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通过邻近连接和定量PCR检测酿酒酵母同源重组中间体

DOI:

10.3791/64240-v

•

07:55 min

•

September 11, 2022

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Diedre Reitz, Jérôme Savocco, Aurèle Piazza, Wolf-Dietrich Heyer³

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

Capítulos

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

Tadução automática

D环捕获（DLC）和D环延伸（DLE）测定利用邻近连接原理与定量PCR一起量化酿酒酵母中诱导双链断裂部位的D环形成，D环延伸和产物形成。

Tags

Keywords: Homologous Recombination D-loop Proximity Ligation Quantitative PCR Saccharomyces Cerevisiae BRCA2 Bloom Syndrome Werner Syndrome UV Cross-linking Spheroplasting Restriction Enzyme Buffer

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