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Simple and cost-effective fluorescent labeling of 5-hydroxymethylcytosine.
- Source :
-
Methods and applications in fluorescence [Methods Appl Fluoresc] 2016 Oct 07; Vol. 4 (4), pp. 044003. Date of Electronic Publication: 2016 Oct 07. - Publication Year :
- 2016
-
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.
Details
- Language :
- English
- ISSN :
- 2050-6120
- Volume :
- 4
- Issue :
- 4
- Database :
- MEDLINE
- Journal :
- Methods and applications in fluorescence
- Publication Type :
- Academic Journal
- Accession number :
- 28192296
- Full Text :
- https://doi.org/10.1088/2050-6120/4/4/044003