Genome-Wide Detection of Meiotic DNA Double-Strand Break Hotspots Using Single-Stranded DNA
The controlled fragmentation of chromosomes by DNA double-strand breaks (DSBs) initiates meiotic recombination, which is essential for meiotic chromosome segregation in most eukaryotes. This chapter describes a straightforward microarray-based approach to measure the genome-wide distribution of meiotic DSBs by detecting the single-stranded DNA (ssDNA) that transiently accumulates at DSB sites during recombination. The protocol outlined here has been optimized to detect meiotic DSBs in Saccharomyces cerevisiae. However, because ssDNA is a universal intermediate of homologous recombination, this method can ostensibly be adapted to discover and analyze programmed or damage-induced DSB hotspots in other organisms whose genome sequence is available.
Key wordsssDNA meiosis double-strand breaks hotspots microarray
We would also like to thank Gerben Vader and Milan de Vries for technical discussions and critical reading of this protocol.
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