Cookies on this website
We use cookies to ensure that we give you the best experience on our website. If you click 'Continue' we'll assume that you are happy to receive all cookies and you won't see this message again. Click 'Find out more' for information on how to change your cookie settings.

We here review primary methods used in quantifying and mapping 5-hydroxymethylcytosine (5hmC), including global quantification, restriction enzyme-based detection, and methods involving DNA-enrichment strategies and the genome-wide sequencing of 5hmC. As discovered in the mammalian genome in 2009, 5hmC, oxidized from 5-methylcytosine (5mC) by ten-eleven translocation (TET) dioxygenases, is increasingly being recognized as a biomarker in biological processes from development to pathogenesis, as its various detection methods have shown. We focus in particular on an ultrasensitive single-molecule imaging technique that can detect and quantify 5hmC from trace samples and thus offer information regarding the distance-based relationship between 5hmC and 5mC when used in combination with fluorescence resonance energy transfer.

Original publication




Journal article



Publication Date





single molecule imaging, epigenetic modification, 5-Methylcytosine, Animals, Antibody Specificity, Base Sequence, Chromatography, High Pressure Liquid, Chromosome Mapping, DNA, DNA Restriction Enzymes, Epigenesis, Genetic, Fluorescence Resonance Energy Transfer, Genetic Markers, Glycosylation, Humans, Mass Spectrometry, Sequence Analysis, DNA, Single Molecule Imaging