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

Catalog No.	Quantity
FERK1501	25 reactions

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

Specifications

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

Frequently Asked Questions (FAQs)

What is the difference between EpiJET 5-hmC Analysis Kit (Cat. No. K1481) and EpiJET 5-hmC and 5-mC Analysis Kit (Cat. No. K1501)?

The new EpiJET 5-hmC and 5-mC Analysis Kit is more universal than the previous EpiJET 5-hmC Analysis Kit because it provides a total assessment of both cytosine modifications - 5-hydroxymethylcytosine (5-hmC) and methylcytosine (5-mC) - within specific DNA loci (for comparison - the current kit allows only 5-hmC analysis). Additionally, all necessary DNA controls do not originate from plasmids widely used in molecular biology laboratories are included into the new kit.

How much DNA can I use for glucosylation and restriction digestion reactions with EpiJET 5-hmC and 5-mC Analysis Kit?

For each glucosylation reaction, a broad range of DNA input can be used from 50 ng to 1 µg.

Do I need an extra step of genomic DNA purification prior to using the sample in EpiJET 5-hmC and 5-mC Analysis Kit?

DNA purity is an important factor for successful digestion by Epi MspI/HpaII. We recommend use of spin column-based kits such as the Thermo Scientific GeneJET Genomic DNA Purification Kit (Cat. No. K0721/2) for genomic DNA purification.

Can I buy T4 B-glucosyltransferase separately from EpiJET 5-hmC and 5-mC Analysis Kit?

Yes, Thermo Scientific T4 B-glucosyltransferase is available as a stand-alone enzyme (Cat. No. EO0831, 500 U).

Safety and Handling

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

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