Quest 5-hmC Detection-Lite Kit w/ Zymo-Spin™ I Columns (Uncapped)

The Quest 5-hmC Detection Kit™ - Lite from Zymo Research allows for sequence specific detection of 5-hydroxymethylcytosine within DNA using a simple and efficient reaction setup. Utilizing a robust and highly specific 5-hmC Glucosyltransferase enzyme, 5-hydroxymethylcytosine in DNA is specifically tagged with a glucose moiety yielding glucosyl-5-hydroxymethylcytosine. Following glucosylation of 5-hydroxymethylcytosine, digestion of DNA with “glucosyl-5-hydroxymethylcytosine sensitive” restriction endonucleases (GSRE) allows for effective differentiation of 5-methylcytosine from 5-hydroxymethylcytosine. Identification of 5-hydroxymethylcytosine in a sequence specific context can then be deduced from the restriction endonuclease recognition sequence. The Quest 5-hmC Detection Kit™ - Lite is adaptable for use with GlaI, MspI, TaqαI, McrBC or any other GSRE. Methods for the setup of convenient and reliable one tube reactions are described for use with GlaI or MspI in the protocol provided.