Buhler Cyril, Shroff Robert, Lichten Michael
Center for Cancer Research, National Cancer Institute, Bethesda, MD, USA.
Methods Mol Biol. 2009;557:143-64. doi: 10.1007/978-1-59745-527-5_10.
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.
DNA双链断裂（DSBs）引发真核生物减数分裂重组。我们描述了两种利用微阵列来确定酿酒酵母中减数分裂DSBs全基因组分布的策略。第一种是染色质免疫沉淀（ChIP）方法，该方法靶向Spo11蛋白，在某些突变背景下，Spo11蛋白会共价连接到DSB末端。第二种方法涉及通过BND纤维素富集在去除Spo11后于DSB位点经末端切除形成的单链DNA（ssDNA）重组中间体。
