The Quest 5-hmC Detection Kit allows for locus-specific detection of 5-hydroxymethylcytosine (5-hmC) using a simple and efficient reaction setup. Traditional methods cannot distinguish 5-hmC from 5-methylcytosine (5-mC). This kit features a robust and highly specific 5-hmC glucosyltransferase enzyme to specifically tag 5-hmC sites, yielding the modified base, glucosyl-5-hydroxymethylcytosine (g-5-hmC). After glucosylation of 5-hmC, digestion of DNA with Msp1 allow 5-hmC to be differentiated from 5-mC. Msp1 can efficiently digest DNA when a cytosine, 5-mC, or 5-hmC is present in their recognition site, but it is sensitive to the presence of g-5-hmC. By exploiting this sensitivity, the 5-hmC level of a specific locus can be interrogated by utilizing various downstream applications (e.g. endpoint PCR, qPCR, Next-Generation sequencing, etc.).