EpiDirect® is a novel qPCR-based method for direct anlysis of DNA methylation status in human samples. Purified DNA is analysed directly without prior bisulfite conversion.
Assays | DNA methylation
Assays | DNA methylation
No pre-treatment | 2 hour run-time
DNA methylation is a epigenetic modification of the cytosine base where a methyl-group is added to the 5th carbon. DNA methylation cannot be analysed directly by PCR using current state-of-the-art methods, that are based on standard DNA chemistries due to lack of discrimination between methylated and unmethylated cytosines. Currently, one of the most applied methods to overcome the issue of analysing DNA methylation with PCR is bisulifte conversion.
Our EpiDirect® qPCR Assay makes it possible to evaluate the methylation status of the MGMT promoter in human cancer biopsies using Real-Time PCR. The analysis does not require prior bisulfite conversion and offers objectivity in measurement, minimum hands-on time, and reduced turn-around time compared to the current state-of-the-art.
PentaBase’s properitary INA® technology is used in the EpiDirect® platform. The INA® works by increasing the p-stacking effect of DNA which increases the affinity for methylated DNA.
The EpiDirect® platform utilizes a special primer design called an EpiPrimer™. The EpiPrimer™ binds strongly to methylated DNA allowing for amplification of the methylated region. This creates a small difference in Ct between the methylated region and a reference gene (DCt). If the DNA is unmethylated, the primer will only bind weakly, creating a large DCt.
The technology behind EpiDirect® is also presented in an article published in Nature Communications in August 2023.
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Use the below links for more information about our first EpiDirect® assay or our INA® technology
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