| Article ID | Journal | Published Year | Pages | File Type |
|---|---|---|---|---|
| 8340747 | Methods | 2015 | 7 Pages |
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 (R2Â >Â 0.99) detection of 5hmC in human brain tissue using the optimised oxBS-450K protocol described here.
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Authors
Sabrina K. Stewart, Tiffany J. Morris, Paul Guilhamon, Harry Bulstrode, Martin Bachman, Shankar Balasubramanian, Stephan Beck,