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The EpiJET 5-hmC Analysis Kit includes two advanced enzymes: T4 BGT and Epi MspI. The T4 BGT is formulated for highly specific, complete, and fast glucosylation of 5-hmC in 15 min. The Epi MspI is an MspI restriction enzyme specially formulated for efficient use in epigenetic studies. It completely digests genomic DNA in 4 hours when cytosine, 5-methylcytosine, or 5-hydroxymethylcytosine is within its recognition sequence, but its activity is completely blocked when 5-hmC is glucosylated. The analysis involves DNA sample glucosylation and subsequent Epi MspI digestion. Restriction digestion products are then analyzed by qPCR with a primer pair flanking CCGG site of interest (see Figure 1 in Supporting Data).