EpiDirect® MGMT Methylation Assay
A bisulfite-free MGMT methylation assay designed for rapid and accurate glioblastoma diagnostics. Using our proprietary INA® chemistry and EpiPrimer™ oligonucleotides, the assay selectively amplifies methylated target DNA directly from unconverted samples.
With as little as 0.5 ng of DNA, EpiDirect® MGMT provides reliable quantification of MGMT promoter methylation in just 2 hours using standard real-time PCR instruments. The assay is compatible with FFPE and fresh-frozen glioblastoma samples and eliminates the need for time-consuming bisulfite conversion.
- Direct qPCR analysis without DNA pretreatment
- High sensitivity: detects methylation levels as low as 2.5%
- CE-IVD certified for clinical use (est. CE IVDR Q4 2025 )
- Fast results in under 2 hours
*Validated on CFX96/CFX Opus 96 (Bio-Rad), QuantStudio™5 (ABI) and BaseTyper™ 48.4 (PentaBase) Real-Time PCR Systems
Product Details
Reference Number 8300
Product Name EpiDirect® MGMT Methylation qPCR Assay
Product Description PCR master mix and oligonucleotide mix for quantification of MGMT promoter methylation, positive and negative control
Format Ready-to-Use
Reactions 48
Intended Use
EpiDirect® MGMT Methylation qPCR Assay is a semi-automated real-time Polymerase Chain Reaction (PCR) assay intended for the direct, semi-quantitative analysis of the methylation status of the human DNA repair enzyme O6-methylguanine-DNA methyltransferase (MGMT) gene promoter.
The assay is intended for analysis of purified genomic DNA (gDNA) samples that without further pre-treatment can be used with real-time PCR systems for the detection and quantification of the methylation status of the MGMT promoter. Samples comprise Formalin-Fixed Paraffin-Embedded (FFPE) and fresh frozen biopsies and can be prepared using automated platforms or manual workflows.
EpiDirect® MGMT Methylation qPCR Assay is intended for use by health professionals or qualified laboratory personnel specifically instructed and trained in the techniques of Real-Time PCR as well as proficient in handling biological samples. Medical interventions based on results from this product requires medical authorization.
Product Specifications
Procedure Real-time PCR-based
Validated Real-Time PCR instruments BaseTyper™ 48.4 Quiet HRM (PentaBase Ref. No. 754), CFX96 and CFX Opus 96 (Bio-Rad) and QuantStudio™ 5 Real-Time PCR Systems
PentaBase technologies EpiPrimer™, HydrolEasy® Probe, and SuPrimer™
Genomic target DMR2 region CpG sites 75-78 in exon 1 of the MGMT promoter
Sample types Solid glioblastoma biopsies including formalin-fixed paraffin-embedded (FFPE) tissue samples
Sample tumor cell percentage At least 20%
Sample DNA extraction methods Relevant commercially avaliable solid biospy/FFPE DNA extraction kits
Sample input 0.1-10 ng/µL of purified DNA directly from the sample that has not been otherwise chemically manipulated
PCR run time Less than 2 hours
Instructions For Use
The latest version of the Instructions For Use can be found at: www.5ba.se/ED001
Publications
A qPCR technology for direct quantification of methylation in untreated DNA
Bendixen et al. – Nature Communications – 2023
Related files
Why Choose EpiDirect® MGMT?
Current methods for evaluating DNA methylation typically rely on sodium bisulfite or other chemical and enzymatic conversion of cytosines. However, variability in conversion efficiency and DNA degradation caused by chemical treatment can lead to false or inconclusive results. In particular, bisulfite treatments often cause biased degradation of unmethylated DNA, which may overestimate methylation levels.
In contrast, the EpiDirect® technology allows for direct quantification of DNA methylation by PCR without any prior chemical or enzymatic treatment. As a result, it eliminates artefacts related to incomplete conversion of cytosines and DNA degradation, while vastly reducing hands-on time and total time to result.
As a result, EpiDirect® MGMT provides accurate, reproducible, and efficient methylation analysis, making it ideal for both research and clinical applications..
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.
