oxBS-450K: a method for analysing hydroxymethylation using 450K BeadChips

Sabrina K Stewart, Tiffany J Morris, Paul Guilhamon, Harry Bulstrode, Martin Bachman, Shankar Balasubramanian, Stephan Beck

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish
Pages (from-to)9-15
Number of pages7
JournalMethods
Volume72
DOIs
Publication statusPublished - 15 Jan 2015

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