Authors:
SK Stewart, TJ Morris, P Guilhamon, H Bulstrode, M Bachman, S Balasubramanian, S Beck
Journal name: 
Methods
Citation info: 
72:9-15
Abstract: 
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.
DOI: 
http://doi.org/10.1016/j.ymeth.2014.08.009
Research group: 
Balasubramanian Group
E-pub date: 
15 Jan 2015