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

Catalog No. FERK1501
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FERK1501 25 reactions
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Catalog No. FERK1501 Supplier Thermo Scientific™ Supplier No. K1501

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 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
TRUSTED_SUSTAINABILITY

Specifications

Compatible Buffer 10X Epi Buffer
Quantity 25 reactions
Product Type 5-hmC and 5-mC Analysis Kit
Product Line EpiJET

Safety and Handling

WARNING: Reproductive Harm - www.P65Warnings.ca.gov

For Research Use Only. Not for use in diagnostic procedures.

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