Meiotic crossover hotspots and coldspots within Arabidopsis disease resistance genes

To study recombination at the fine-scale we used high-throughput sequencing to analyse >1,000 crossovers within the RAC1 R gene hotspot. This revealed focused intragenic crossovers, overlapping exons encoding the TIR, NBS and LRR domains (RAC1 pollen typing sequencing). To analyse chromatin structure we performed micrococcal nuclease digestion of wild type (Col-0) chromatin and gel purified the resulting ~150 bp mononucleosomal DNA band. This DNA was used to generate a library and paired-end sequencing performed (MNase-seq)

为在精细尺度下研究重组现象，我们采用高通量测序技术，对RAC1 R基因热点区域内的超过1000个重组交换位点进行分析。该分析揭示了集中分布的基因内重组交换事件，这些交换位点与编码TIR、NBS及LRR结构域的外显子区域重叠（RAC1花粉分型测序）。为分析染色质结构，我们对野生型（Col-0）染色质进行微球菌核酸酶酶切，并通过凝胶纯化得到约150 bp的单核小体DNA条带。以此DNA构建测序文库并开展双端测序（MNase-seq）