Division of Epigenomics and Development, Medical Institute of Bioregulation, Kyushu University, Fukuoka, Japan.
Methods Mol Biol. 2022;2509:233-250. doi: 10.1007/978-1-0716-2380-0_14.
Whole genome bisulfite sequencing (WGBS) is a high-throughput DNA sequencing-based technique that is used to determine genome-wide DNA methylation patterns at base resolution. Library construction by post-bisulfite adaptor tagging (PBAT ) extends the application of WGBS to several hundred cells and minimizes the required number of library amplification cycles. We herein describe a PBAT protocol to prepare WGBS libraries from 200 cells and introduce the outline of a downstream bioinformatic analysis. The prepared library can typically generate 800 million sequencing reads, which is sufficient to cover the human and mouse genomes approximately 15 times, using the Illumina NovaSeq 6000 sequencing system.
全基因组亚硫酸氢盐测序（WGBS）是一种高通量的基于 DNA 测序的技术，用于在碱基分辨率上确定全基因组 DNA 甲基化模式。通过 post-bisulfite adaptor tagging（PBAT）进行文库构建，将 WGBS 的应用扩展到几百个细胞，并将文库扩增循环所需的数量最小化。本文描述了一种从 200 个细胞制备 WGBS 文库的 PBAT 方案，并介绍了下游生物信息学分析的概要。使用 Illumina NovaSeq 6000 测序系统，通常可以生成 8 亿个测序读段，足以覆盖人类和小鼠基因组约 15 倍。
