Genome-wide mapping of meiotic DNA double-strand breaks in Saccharomyces cerevisiae.
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.
Center for Cancer Research, National Cancer Institute, Bethesda, MD, USA.
Cyril
C
Buhler
Robert
R
Shroff
Michael
M
Lichten
D018929:Cell Culture Techniques / Q000379:methods; D047369:Chromatin Immunoprecipitation / Q000379:methods; D002874:Chromosome Mapping / Q000379:methods*; D053903:DNA Breaks, Double-Stranded; D020869:Gene Expression Profiling / Q000379:methods; D016681:Genome, Fungal; D008540:Meiosis / Q000235:genetics*; D008954:Models, Biological; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D016133:Polymerase Chain Reaction / Q000379:methods; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000254:growth & development