Thermo Scientific™ EpiJET™ DNA Methylation Analysis Kit (TaqI/HpyF30I)

5-Methylcytosine is a prominent epigenetic DNA modification which plays an important role in up- and down-regulation of gene expression. In mammalian cells 5-methylcytosines are mainly found in 5'-CpG-3' dinucleotides and are enriched in CpG islands, the GC-rich regions most often located in gene promoters. The MspI/HpaII enzyme pair with 5'-CCGG-3' recognition sequence has multiple targets in such regions and is routinely used for cytosine methylation analysis. Outside the CpG islands the CCGG motifs are much less abundant and limit the usability of the aforementioned enzyme pair.

The Thermo Scientific™ EpiJET DNA Methylation Analysis Kit (HpyF30I/TaqI) uses another pair of restriction enzymes to analyze the DNA methylation status in gene bodies and other low-GC content genomic regions. HpyF30I and TaqI are isoschizomers with TCGA specificity, when cytosine in the 5'-TCGA-3' tetranucleotide is methylated, the cleavage with Epi HpyF30I is blocked, but cleavage with Epi TaqI is not affected.

The Epi HpyF30I and Epi TaqI enzymes are specially formulated for the epigenetic studies to complete genomic DNA digestion in 1 hour.

5-Methylcytosine is a prominent epigenetic DNA modification which plays an important role in up- and down-regulation of gene expression. In mammalian cells 5-methylcytosines are mainly found in 5'-CpG-3' dinucleotides and are enriched in CpG islands, the GC-rich regions most often located in gene promoters. MspI/HpaII enzyme pair with 5'-CCGG-3' recognition sequence has multiple targets in such regions and is routinely used for cytosine methylation analysis. Outside the CpG islands the CCGG motifs are much less abundant and limit the usability of the aforementioned enzyme pair.

The Thermo Scientific EpiJET DNA Methylation Analysis Kit (HpyF30I/TaqI) uses another pair of restriction enzymes to analyze the DNA methylation status in gene bodies and other low-GC content genomic regions. HpyF30I and TaqI are isoschizomers with TCGA specificity, when cytosine in the 5'-TCGA-3' tetranucleotide is methylated, the cleavage with Epi HpyF30I is blocked, but cleavage with Epi TaqI is not affected.

The Epi HpyF30I and Epi TaqI enzymes are specially formulated for the epigenetic studies to complete genomic DNA digestion in 1 hour.

Highlights

• Unique - novel tool for DNA methylation analysis in 5' TCGA 3' sequences
• Efficient - enzymes specifically formulated for genomic DNA methylation analysis
• Robust - reproducible data, correlating with other analysis methods
• Fast procedure - complete digestion of genomic DNA in 1h
  Applications
• CpG methylation analysis at the 5'-TCGA-3' loci
  Includes
• EpiJET HpyF30I
• Epi TaqI
• 10X Epi Buffer 2
• Control DNA Unmethylated (pUC19/SmaI) (0.5 μg/μL)
• Control DNA CpG Methylated (mpUC19/SmaI) (0.5 μg/μL)

Hazardous: No
Quality Control:

• The kit is tested in Epi HpyF30I and Epi TaqI digestion of human blood genomic DNA, spiked with methylated and unmethylated pUC19DNA/SmaI.
• Unmethylated plasmid DNA and genomic DNA are completely cleaved in one hour with both enzymes.
• Methylated plasmid DNA and genomic DNA is not cleaved by Epi HpyF30I, but cleaved with Epi TaqI.