EpiJET DNA Methylation Analysis Kit (TaqI/HpyF30I)

Detailní popis

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)

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)