Journal
METHODS
Volume 72, Issue -, Pages 9-15Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ymeth.2014.08.009
Keywords
Hydroxymethylation; DNA methylation; 450K BeadChip; Oxidation; Bisulfite conversion; Epigenetics
Funding
- MRC CASE studentship [G1000411]
- Wellcome Trust [99148]
- Royal Society Wolfson Research Merit Award [WM100023]
- EU-FP7 project IDEAL [259679]
- EU-FP7 project EPIGENESYS [257082]
- EU-FP7 project BLUEPRINT [282510]
- Cancer Research UK [11961] Funding Source: researchfish
- Medical Research Council [973748] Funding Source: researchfish
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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-hydroxymethyl-cytosine (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. (C) 2014 The Authors. Published by Elsevier Inc.
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