DMMC Course: UNRAVELLING CHROMATIN & THE ROLE OF EPIGENETICS IN DISEASE

1115-1140 Tuesday 25 April 2006. UCD Conway Institute Lecture Theatre

Assessing DNA methylation in the laboratory
Antoinette Perry (Institute of Molecular Medicine, TCD)

DNA methylation is recognised as having a significant role in a variety of physiological and pathological processes and particularly in human carcinogenesis. Many new technologies have been developed over the last decade to examine promoter hypermethylation and to dissect complex epigenetic patterns. These can broadly be considered as either candidate gene or genome-wide approaches. The most widely employed techniques such as DNA sequencing and methylation specific PCR rely on bisulfite conversion of genomic DNA, which permits discrimination between methylated and unmethylated DNA based on synthetic base changes. Demethylating agent 5-azacytidine is routinely used to examine the effects of promoter hypermethylation on gene expression. 5-azacytidine is now being used in combination with large-scale methylation arrays to identify thousands of targets of hypermethylation in a single experiment. This talk will provide an overview of current methylation technologies, with specific attention given to a handful of methods, common problems encountered and important controls that would be helpful for anybody interested in working on DNA methylation.