Genome-wide mapping of meiotic DNA double-strand breaks in Saccharomyces cerevisiae

Methods Mol Biol. 2009:557:143-64. doi: 10.1007/978-1-59745-527-5_10.

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.

Publication types

  • Review

MeSH terms

  • Cell Culture Techniques / methods
  • Chromatin Immunoprecipitation / methods
  • Chromosome Mapping / methods*
  • DNA Breaks, Double-Stranded*
  • Gene Expression Profiling / methods
  • Genome, Fungal
  • Meiosis / genetics*
  • Models, Biological
  • Oligonucleotide Array Sequence Analysis / methods
  • Polymerase Chain Reaction / methods
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae / growth & development