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

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Stewart, Sabrina K., Morris, Tiffany J., Guilhamon, Paul, Bulstrode, Harry, Bachman, Martin, Balasubramanian, Shankar, Beck, Stephan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Academic Press 2015
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4304834/
https://www.ncbi.nlm.nih.gov/pubmed/25175075
http://dx.doi.org/10.1016/j.ymeth.2014.08.009
Descripción
Sumario: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.

Ejemplares similares