Detection of hydroxymethylcytosine bases

Abstract

Provided herein is a method of labeling the epigenetic modification 5-hydroxymethyl-cytosine along a DNA molecule that involves attaching a 5-hydroxymethyl-cytosine specific labeling agent to the DNA molecule, which is effected by enzymatically synthesizing UDP-6-N3-Glucose by subjecting 6-azidoglucose to an enzymatic catalysis by N-acetylhexoseamine 1-kinase (NahK) in the presence of ATP to thereby obtain a phosphorylated 6-azidoglucose, followed by subjecting the phosphorylated 6-azidoglucose to enzymatic catalysis by uridyltransferase (GlmU) in the presence of UTP to thereby obtain UDP-6-N3-Glucose, followed by incubating the DNA molecule with UDP-6-N3-Glucose and ß-glucosyltransferase, to thereby obtain a 5-hydroxymethyl-cytosin e glycosylated by a 6-azidoglucose residue in the DNA molecule, and reacting the modified DNA molecule with a labeling agent which is chemically compatible with the azide group of the 6-azidoglucose residue.

Dependencies are inferred from the claim text.