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EpiJET DNA Methylation Analysis Kit (MspI/HpaII)


The EpiJET DNA Methylation Analysis Kit (MspI/HpaII) uses MspI and HpaII for fast and efficient analysis of the DNA methylation status at a specific locus.
  
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5-Methylcytosine is a prominent epigenetic DNA modification which plays an important role in regulation of gene expression.

The Thermo Scientific EpiJET DNA Methylation Analysis Kit (MspI/HpaII) uses the MspI and HpaII restriction enzymes to analyze DNA methylation status at a specific locus. Epi MspI and Epi HpaII are isoschizomers with differing sensitivities to CpG methylation. When the internal CpG in the 5’-CCGG-3’ tetranucleotide sequence is methylated, cleavage with Epi HpaII is blocked, but cleavage with Epi MspI is not affected.

The Epi MspI and Epi HpaII enzymes are specially formulated for epigenetics studies to complete genomic DNA digestion in 1 hour.

Highlights

  • Fast – complete digestion of genomic DNA in 1 hour at 37°C
  • Efficient – specially formulated enzymes for epigenetic studies of DNA methylation status

Applications

  • CpG methylation analysis at 5’-CCGG-3’ loci

Includes

  • Epi Mspl: 200 µL
  • Epi HpaII: 200 µL
  • 10X EpiBuffer: 1 mL
  • Control pUC19/SmaI DNA Unmethylated (0.5 µg/µL): 20 µL
  • Control pUC19/SmaI DNA CpG Methylated (0.5 µg/µL): 20 µL

Note

DNA sample quality is important for efficient digestion by restriction endonucleases or downstream applications. We recommend use of spin column based kits, such as the Thermo Scientific GeneJET Genomic DNA Purification Kit (#K0721, #K0722).

  
Storage Condition-20 C
HazardousNo
10X EpiBuffer Composition1 mg/mL BSA, 330 mM Tris-acetate (pH 7.9), 1 mM Acetic acid, 660 mM Potassium acetate
Quality ControlFunctional Testing
  • The kit is functionally tested in Epi MspI and Epi HpaII digestion of human blood genomic DNA, spiked with methylated and unmethylated pUC19DNA/SmaI.
  • Unmethylated plasmid and genomic DNA are completely cleaved in one hour with both enzymes.
  • Methylated plasmid and genomic DNA are not cleaved by Epi HpaII, but cleaved by Epi MspI.
Parallel digestion of methylated and unmethylated DNA with the restriction enzyme pair (MspI/HpaII)

Parallel digestion of methylated and unmethylated DNA with the restriction enzyme pair (MspI/HpaII)

Parallel digestion of methylated and unmethylated DNA with the restriction enzyme pair (MspI/HpaII)

Parallel digestion of methylated and unmethylated DNA with the restriction enzyme pair (MspI/HpaII) having different sensitivity to methylation.

Methylated and unmethylated linearized plasmid DNA (pUC19/SmaI) were mixed with human blood genomic DNA and digested in parallel with methylation-sensitive Epi HpaII restriction enzyme and methylation insensitive isoschizomer Epi MspI for 1 hour at 37°C. Methylated plasmid DNA and naturally methylated gDNA were protected from cleavage with Epi HpaII, but were cleaved with insensitive Epi MspI. Unmethylated plasmid DNA was cleaved with both enzymes.

1, 4 – undigested DNA (control)

2, 5 – after digestion with Epi HpaII

3, 6 – after digestion with Epi MspI