Stewart, SK;
Morris, TJ;
Guilhamon, P;
Bulstrode, H;
Bachman, M;
Balasubramanian, S;
Beck, S;
(2014)
oxBS-450K: A method for analysing hydroxymethylation using 450K BeadChips.
Methods
, 72
pp. 9-15.
10.1016/j.ymeth.2014.08.009.
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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.
| Type: | Article |
|---|---|
| Title: | oxBS-450K: A method for analysing hydroxymethylation using 450K BeadChips |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1016/j.ymeth.2014.08.009 |
| Publisher version: | http://dx.doi.org/10.1016/j.ymeth.2014.08.009 |
| Language: | English |
| Additional information: | Copyright © 2014 The Authors. Published by Elsevier Inc. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/3.0/). |
| Keywords: | 450K BeadChip, Bisulfite conversion, DNA methylation, Epigenetics, Hydroxymethylation, Oxidation |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Cancer Institute UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Cancer Institute > Research Department of Cancer Bio |
| URI: | https://discovery-pp.ucl.ac.uk/id/eprint/1447467 |
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