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

On demand
Order number : K1501
(Ex-work price)
$ 394

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

 

Themo Scientific 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.

 

Features

  • Efficient and reliable—accurate quantification of 5-hmC and 5-mC
  • Robust—broad range (from 0.2 µg to 4 µg) of input DNA
  • Fast—procedure completed in 5 hours, no overnight incubation.

 

Contents

  • T4 ß – glucosyltransferase, 5 U/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
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