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Meiosis

Volume 557 of the series Methods in Molecular Biology pp 143-164

Date:

Genome-Wide Mapping of Meiotic DNA Double-Strand Breaks in Saccharomyces cerevisiae

  • Cyril BuhlerAffiliated withCenter for Cancer Research, National Cancer Institute
  • , Robert ShroffAffiliated withCenter for Cancer Research, National Cancer Institute
  • , Michael LichtenAffiliated withLaboratory of Biochemistry and Molecular Biology, Center for Cancer Research, National Cancer Institute

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Abstract

DNA double-strand breaks (DSBs) initiate meiotic recombination in eukaryotes. We describe two strategies that use microarrays to determine the genome-wide distribution of meiotic DSBs in the yeast Saccharomyces cerevisiae. The first is a chromatin immunoprecipitation (ChIP) approach that targets the Spo11 protein, which remains covalently attached to DSB ends in certain mutant backgrounds. The second approach involves BND cellulose enrichment of the single-strand DNA (ssDNA) recombination intermediate formed by end-resection at DSB sites following Spo11 removal.

Key words

Double-strand break Spo11 DMC1 rad50S chromatin immunoprecipitation single-strand DNA BND cellulose microarray background normalization quantitative PCR