Monitoring Gene Conversion in Budding Yeast by Southern Blot Analysis
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By using an inducible site-specific double-strand break (DSB) in budding yeast, it is possible to monitor—in real time—the repair of the break by homologous recombination. A method is described using an ectopic homologous donor sequence to repair an HO endonuclease-induced DSB. These gene conversion events can occur with or without crossing-over, the products of which are distinguished as different-sized restriction endonuclease fragments. The method of Southern blotting is described in detail.
Key wordsSaccharomyces cerevisiae Budding yeast Double-strand break repair Gene conversion Crossing-over Southern blot
- 10.Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, pp 382–386Google Scholar
- 12.Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–17Google Scholar