Systematic evaluation of library preparation methods and sequencing platforms for high-throughput whole genome bisulfite sequencing [WGBS]

We systematically evaluated alternate strategies for WGBS studies, taking into account opportunites around library preparation methods, sequencing platforms and analysis pipeline optimization. We also assessed the performance and precision of the WGBS method relative to the methylation arrays, in an effort to provide data-driven recommendations for future WGBS studies, in particularly with respect to minimum coverage. Overall design: Bisulfite-converted DNA for 8 human samples were hybridised to the Illumina Infinium 450k Human Methylation Beadchip and Illumina Infinium HumanMethylation850 BeadChip. Whole genome bisulphite sequencing libraries for 8 human samples were prepared by different methods with low DNA input requirement and sequenced by Illumina NovaSeq and HiSeq X10 platforms.

本研究系统评估了全基因组亚硫酸氢盐测序（Whole Genome Bisulfite Sequencing，WGBS）研究中的多种替代策略，综合考量了建库方法、测序平台与分析流程优化等方向的可行方案。同时，本研究对比甲基化芯片，评估了WGBS方法的性能与精确度，以期为未来的WGBS研究提供数据驱动的建议，尤其针对最低测序覆盖度这一维度。
整体实验设计：将8个人类样本的亚硫酸氢盐转化DNA分别与Illumina Infinium 450K人类甲基化芯片及Illumina Infinium HumanMethylation850芯片进行杂交；采用不同低起始量建库方法制备8个人类样本的全基因组亚硫酸氢盐测序文库，并通过Illumina NovaSeq与HiSeq X10测序平台完成测序。