Homo sapiens Epigenomics. Homo sapiens

We sought to develop a method that can partition bisulfite treated DNA into methylated and unmethylated fractions. The method, Streptavidin bisulfite ligand methylation enrichment (SuBLiME) involves specific labelling with a biotin-labelled nucleotide ligand at, or opposite, the site of a methylated cytosine in a bisulfite-converted nucleic acid and then subsequent affinity capture using streptavidin-coupled magnetic beads. This method is highly adaptable and can be combined with deep sequencing library generation and/or genomic complexity reduction. In this pilot study we enriched methylated DNA from the Csp6I cut complexity-reduced genomes of the colorectal cancer cell lines HCT-116, HT-29 and SW-480 and normal blood leukocytes with the aim of discovering colorectal cancer biomarkers.

本研究旨在开发一种可将亚硫酸氢盐处理的DNA划分为甲基化与非甲基化组分的方法。该方法为链霉亲和素-亚硫酸氢盐配体甲基化富集（Streptavidin bisulfite ligand methylation enrichment, SuBLiME），其操作流程为：在亚硫酸氢盐转化的核酸中，于甲基化胞嘧啶位点或其互补位点使用生物素标记的核苷酸配体进行特异性标记，随后借助链霉亲和素偶联磁珠完成后续亲和捕获。该方法适配性极强，可与深度测序文库构建及/或基因组复杂度降低技术联合使用。在本预实验研究中，我们对结直肠癌细胞系HCT-116、HT-29、SW-480以及正常血液白细胞经Csp6I酶切得到的复杂度降低基因组中的甲基化DNA进行了富集，以期发现结直肠癌生物标志物。