Li Qing, Hermanson Peter J, Springer Nathan M
National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan, Hubei, 430070, China.
Department of Plant Biology, Microbial and Plant Genomics Institute, University of Minnesota, Saint Paul, MN, 55108, USA.
Methods Mol Biol. 2018;1676:185-196. doi: 10.1007/978-1-4939-7315-6_11.
DNA methylation plays an important role in the regulation of the expression of transposons and genes. Various methods have been developed to assay DNA methylation levels. Bisulfite sequencing is considered to be the "gold standard" for single-base resolution measurement of DNA methylation levels. Coupled with next-generation sequencing, whole-genome bisulfite sequencing (WGBS) allows DNA methylation to be evaluated at a genome-wide scale. Here, we described a protocol for WGBS in plant species with large genomes. This protocol has been successfully applied to assay genome-wide DNA methylation levels in maize and barley. This protocol has also been successfully coupled with sequence capture technology to assay DNA methylation levels in a targeted set of genomic regions.
DNA甲基化在转座子和基因表达调控中发挥着重要作用。人们已经开发出多种方法来检测DNA甲基化水平。亚硫酸氢盐测序被认为是单碱基分辨率测量DNA甲基化水平的“金标准”。结合下一代测序技术，全基因组亚硫酸氢盐测序（WGBS）能够在全基因组范围内评估DNA甲基化情况。在此，我们描述了一种适用于大基因组植物物种的WGBS方案。该方案已成功应用于检测玉米和大麦全基因组的DNA甲基化水平。该方案还成功地与序列捕获技术相结合，用于检测一组目标基因组区域的DNA甲基化水平。
