This technology is based on a simple restriction digest followed by quantitative real-time PCR detection of the genes of interest. The system is reliable, reproducible and easy to adapt for high-throughput screening, making it an ideal tool for DNA methylation profiling and biomarker discovery.

DNA methylation plays an important role in gene expression and it occurs almost exclusively in the context of CpG dinucleotides in the form of a covalent attachment of a methyl residue to the cytosine residue. CpG islands (CGIs) are regions with an elevated GC content and a high frequency of CpG dinucleotides which overlap the promoter region of 60-70% of all human genes. Hyper-methylation of CpG islands at gene promoters is mostly associated with gene silencing.

The EpiTect Methyl qPCR Array System relies on the differential cleavage of target sequences by two different restriction endonucleases whose activities require either the presence or absence of methylated cytosines in their respective recognition sequences. As real-time PCR quantifies the relative amount of DNA remaining after each enzyme digestion, the methylation status of individual genes and the methylation profile across a gene panel are reliably and easily calculated. The high yield of DNA from the restriction digests and PCR amplification allow the analysis of smaller, more heterogeneous samples.

For confirmation of data obtained by the EpiTect Methyl qPCR system, other technologies such as methylation-specific PCR (MSP), MethyLight, high resolution melting (HRM), or Pyrosequencing can be used. These technologies are based on QIAGEN’s proprietary bisulfite conversion technology. Visit the Epigenetics overview page for more information.

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