Chen Yun-Ru, Yu Sheng, Zhong Silin
School of Life Sciences, The Chinese University of Hong Kong, Shatin, Hong Kong, China.
Methods Mol Biol. 2018;1675:31-43. doi: 10.1007/978-1-4939-7318-7_2.
DNA cytosine methylation is one of the most abundant epigenetic marks found in the plant nuclear genome. Bisulfite sequencing (BS-Seq) is the method of choice for profiling DNA cytosine methylation genome-wide at a single nucleotide resolution. The basis of this technique is that the unmethylated cytosine can be deaminated to uracil by sodium bisulfite, while the methylated cytosine is resistant to the treatment. By deep sequencing of the bisulfite converted genomic DNA, the methylation level of each mappable cytosine position in the genome could be measured. In this chapter, we present a detailed 2-day protocol for performing a BS-Seq experiment and a simple bioinformatic workflow for wet lab biologists to visualize the methylation data.
DNA胞嘧啶甲基化是植物核基因组中最丰富的表观遗传标记之一。亚硫酸氢盐测序（BS-Seq）是在单核苷酸分辨率下对全基因组DNA胞嘧啶甲基化进行分析的首选方法。该技术的基础是，未甲基化的胞嘧啶可被亚硫酸氢钠脱氨基转化为尿嘧啶，而甲基化的胞嘧啶则对该处理具有抗性。通过对亚硫酸氢盐转化后的基因组DNA进行深度测序，可以测量基因组中每个可映射胞嘧啶位置的甲基化水平。在本章中，我们提供了一个详细的为期两天的BS-Seq实验方案，以及一个简单的生物信息学工作流程，供湿实验室生物学家可视化甲基化数据。
