Monitoring Gene Conversion in Budding Yeast by Southern Blot Analysis

Abstract

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.