Quest 5-hmC detection kits are for sequence- and locus-specific detection of 5-hydroxymethylcytosine (5-hmC) within DNA. Featured is an optimized 5-hmC glucosyltransferase enzyme that specifically adds glucose to 5-hmC in DNA. Unique endonucleases are utilized to distinguish 5-hmC in the glucose-modified DNA. The Quest 5-hmC detection kit includes all of the enzymes, co-factors, controls, and DNA cleanup technologies necessary for consolidated and efficient 5-hmC detection. DNA modified with the Quest products is useful for qPCR or precise mapping applications facilitated by next-gen sequencing platforms.