title: Simple and cost-effective fluorescent labeling of 5-hydroxymethylcytosine.
abstract: The nucleobase 5-hydroxymethylcytosine (5-hmC), a modified form of cytosine, is an important epigenetic mark related to regulation of gene expression. 5-hmC levels are highly dynamic during early development and are modulated during the progression of neurodegenerative disease and cancer. We describe a spectroscopic method for the global quantification of 5-hmC in genomic DNA. This method relies on the enzymatic glucosylation of 5-hmC, followed by a glucose oxidation step that results in the formation of aldehyde moieties that are covalently linked to a fluorescent reporter by oxime ligation. The fluorescence intensity of the labeled sample is directly proportional to its 5-hmC content. We show that this simple and cost-effective technique is suitable for quantification of 5-hmC content in different mouse tissues.
authors.affiliation: These authors contributed equally to this paper.
authors.forename: Tamar
authors.initials: T
authors.lastname: Shahal
authors.forename: Ori
authors.initials: O
authors.lastname: Green
authors.forename: Uri
authors.initials: U
authors.lastname: Hananel
authors.forename: Yael
authors.initials: Y
authors.lastname: Michaeli
authors.forename: Doron
authors.initials: D
authors.lastname: Shabat
authors.forename: Yuval
authors.initials: Y
authors.lastname: Ebenstein