Thermo Scientific™ EpiJET 5-hmC and 5-mC Analysis Kit

Description

• Description The EpiJET 5-hmC and 5-mC Analysis Kit includes three advanced enzymes: T4 phage β-glucosyltransferase (T4 BGT), Epi MspI and Epi HpaII
• The Epi MspI and Epi HpaII are isoschisomers with differing sensitivity to internal C methylation of the CCGG site
• Epi MspI completely digests genomic DNA when cytosine, 5-methylcytosine (5-mC) or 5-hydroxymethylcytosine (5-hmC) is within its recognition sequence, but its activity is completely blocked when 5-hmC is glucosylated
• Epi HpaII cleaves the unmodified CCGG site only
• The kit takes advantage of the T4 BGT ability to specifically modify 5-hmC residues by adding glucose moiety to 5-hmC and the sensitivity of restriction enzyme Epi MspI to glucosylated 5-hmC
• The T4 BGT is formulated for highly specific, complete and fast glucosylation of 5-hmC in 15 minutes
• The analysis involves DNA sample glucosylation and subsequent restriction digestion with Epi MspI and Epi HpaII
• Restriction digestion products are then analyzed by qPCR with a primer pair flanking CCGG site of interest
• Highlights Efficient and reliable - accurate quantification of 5-hmC and 5-mC Robust - broad range (from 0.2μg to 4μg) of input DNA Fast - procedure completed in 5 hours, no overnight incubation Applications 5-hmC and 5-mC analysis and quantification in CCGG sequences Includes T4 β - glucosyltransferase, 5U/μL 10X Epi Buffer 10X UDP–glucose Epi MspI Epi HpaII 5-hmC Control DNA 5-mC Control DNA Unmodified Control DNA Control primer 1 Control primer 2 Water, nuclease-free Detailed Protocol