EpiJET 5-hmC Analysis Kit

The Thermo Scientific EpiJET 5-hmC Analysis Kit is an efficient system for detection of 5-hydroxymethylcytosine (5-hmC) within specific DNA loci. The kit takes advantage of the T4 phage β-glucosyltransferase (T4 BGT) ability to specifically modify 5-hmC residues by adding glucose moiety to 5-hmC and the sensitivity of restriction enzyme MspI to glucosylated 5-hmC.

Features

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).

Highlights

• Complete and specific 5-hmC conversion to glucosylated 5-hmC in 15 min
• Discrimination of 5-hmC in CCGG sequences by Epi MspI digestion in 4 hours
• Quantification of 5-hmC percentage in CCGG sequences by qPCR

Applications

• 5-hmC analysis and quantification in CCGG sequences

Includes

• T4 β – glucosyltransferase, 5 U/µL
• 10X Epi Buffer
• 10X UDP–glucose
• Epi MspI
• 5-hmC Control DNA
• Control primer 1
• Control primer 2
• Water, nuclease-free 
• Detailed Protocol