oxBS-450K：一种使用450K基因芯片分析羟甲基化的方法。
oxBS-450K: a method for analysing hydroxymethylation using 450K BeadChips.
作者信息
Stewart Sabrina K, Morris Tiffany J, Guilhamon Paul, Bulstrode Harry, Bachman Martin, Balasubramanian Shankar, Beck Stephan
机构信息
Department of Cancer Biology, UCL Cancer Institute, University College London, London WC1E 6BT, UK.
Scottish Centre for Regenerative Medicine, Edinburgh EH16 4UU, UK.
出版信息
Methods. 2015 Jan 15;72:9-15. doi: 10.1016/j.ymeth.2014.08.009. Epub 2014 Aug 28.
DNA methylation analysis has become an integral part of biomedical research. For high-throughput applications such as epigenome-wide association studies, the Infinium HumanMethylation450 (450K) BeadChip is currently the platform of choice. However, BeadChip processing relies on traditional bisulfite (BS) based protocols which cannot discriminate between 5-methylcytosine (5mC) and 5-hydroxymethylcytosine (5hmC). Here, we report the adaptation of the recently developed oxidative bisulfite (oxBS) chemistry to specifically detect both 5mC and 5hmC in a single workflow using 450K BeadChips, termed oxBS-450K. Supported by validation using mass spectrometry and pyrosequencing, we demonstrate reproducible (R(2)>0.99) detection of 5hmC in human brain tissue using the optimised oxBS-450K protocol described here.
DNA甲基化分析已成为生物医学研究不可或缺的一部分。对于诸如全表观基因组关联研究等高通量应用，Infinium HumanMethylation450（450K）芯片目前是首选平台。然而，芯片处理依赖于基于传统亚硫酸氢盐（BS）的方案，该方案无法区分5-甲基胞嘧啶（5mC）和5-羟甲基胞嘧啶（5hmC）。在此，我们报告了对最近开发的氧化亚硫酸氢盐（oxBS）化学方法的改进，以使用450K芯片在单个工作流程中特异性检测5mC和5hmC，称为oxBS-450K。在此处所述的优化oxBS-450K方案的支持下，我们通过质谱和焦磷酸测序验证，证明在人脑组织中可重复（R(2)>0.99）检测5hmC。
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