After decades of fine-tuning the ability to analyze the nucleic acid sequences of DNA, scientists are growing increasingly interested in detecting and understanding the methylation marks that sit on top of that DNA. From locus-specific methylation to genome-wide methylome profiles, this layer of information about gene regulation and expression is very important to studies of disease — especially cancer.
At Quantabio, we often speak with our customers to learn more about the specific assays and kits they’re using so we can provide the best support possible. Recently, we’ve been hearing a lot about EpiMelt kits from MethylDetect, which are based on methylation-sensitive high-resolution melting (MS-HRM) technology. Scientists are turning to these kits for their unique primer design which, in combination with a specific annealing temperature, ensures a robust and highly sensitive DNA methylation assessment.
We evaluated various EpiMelt kits using the Quantabio Q-qPCR platform. In our hands, this generated precise and sensitive results for determining the methylation status of target DNA, particularly from formalin-fixed, paraffin-embedded (FFPE) tissue samples. In a new application note, we describe how we used five gene-specific EpiMelt Kits targeting the biomarkers BRCA1, MLH1, and MGMT, as well as BRCA2 and APC, and analyzed them using the Quantabio Q Instrument.
Each EpiMelt Kit contains three ready-to-use controls: one methylated, one un-methylated, and one for assay calibration. The assay calibration control is included to ensure a unique assay sensitivity. The Quantabio Q-qPCR platform and its user-friendly software allow an easy and clear DNA methylation level assessment.
Our evaluation demonstrated the high sensitivity of the EpiMelt assays, with easily distinguishable differences between the assay calibration control and the 0% methylated control. We analyzed breast cancer samples and were able to clearly identify which were unmethylated and which had low-level methylation. The amplification was robust and reproducible for all three replicates of the five EpiMelt assays, with low Ct values and steep curve slopes before reaching the plateau. The MS-HRM results were uniform and presented a high assay sensitivity.
This approach is quite useful since disease-specific methylation changes can be utilized as biomarkers for disease predisposition screening, early disease detection, personalization of treatment, and post-treatment surveillance. In cancer, for instance, cells are typically hypomethylated genome-wide, while specific hypermethylation is observed in a subset of CpG islands at transcriptional regulatory elements of tumor suppressor genes. The methylation status at specific genes can have distinct consequences for the disease course, as increasing research shows that locus-specific methylation changes can be utilized as biomarkers at all stages of disease development.
To learn more, please check out the full application note: “High-Sensitivity DNA Methylation Analysis Using EpiMelt Kits on the Quantabio Q-qPCR Instrument.”
2min
Oct 31, 2025
