DNA methylation (5mC) and DNA 5-hydroxy methylation (5hmC) analysis in WT murine ES cells

DNA methylation in murine WT ES cells was investigated by MBD domain mediated pull down of methylated DNA followed by chip analysis. For examination of 5hmC, hydroxymethylated DNA was enriched by Click-chemistry mediated pull down. Because of the lower amount of 5hmC, two different approached were adopted. The first approach was based on direct labeling of 5hmC DNA, and in the second method DNA was labeled after amplification with vitro transcription (IVT) . Microarray based hybridization (chip) analysis of epigenetic modifications, namely, 5mC and 5hmC for murine WT ES cells.