EpiDirect® Methylation
qPCR Assays
EpiDirect® is a revolutionising conversion-free approach to evaluate tumor DNA methylation status by real-time PCR. EpiDirect® Methylation Assays are able to directly measure DNA methylation status of purified DNA without the need for prior bisulfite treatment.
Eliminate uncertainties and time spent on bisulfite conversion
Bisulfite conversion is a time and resurse consuming process that can easily lead to overestimation of the methylation frequency. The EpiDirect® approach to assessing DNA methylation status bypasses the bisulfite conversion step and provides results in less than two hours using standard real-time PCR instrumentation.
The EpiDirect® Methylation Assays family targets include the MGMT promoter for use with glioblastoma tumor samples and the MLH1 promoter for use with MSI colorectal and endometrial cancer samples.
How The EpiDirect® Technology Works
The technology relies on a unique DNA analogue chemistry called intercalating pseudo-nucleotides (IPN). These special oligonucleotides, also known as intercalating nucleic acids (INAs®), bind more strongly and specifically to target DNA than standard oligonucleotides.
Central to the system is the EpiPrimer™, which is designed to selectively amplify methylated DNA. Each EpiPrimer™ has three key parts:
- Anchor sequence: Binds to the methylated DNA region.
- Starter sequence: Initiates replication of the methylated target.
- Loop sequence: Supports priming of the remaining DNA during PCR cycles.
Together, the anchor and starter sequences drive initial replication, while the loop and starter sequences handle the later PCR cycles. As a result, the assay achieves accurate and efficient amplification without chemical treatments.
Moreover, this approach reduces errors, saves time, and is easier to use than traditional bisulfite-based methods. In addition, it works on standard PCR instruments, making it practical for both research and clinical labs.
