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| Product Name | Methylation Probe qPCR Master Mix |
| Catalog No. | RESK-0113 |
| Description | This product is a high-performance qPCR reagent specifically designed for methylation detection. Carefully engineered and optimized, it contains a specially processed Taq DNA polymerase, Mg2+, dNTPs, PCR stabilizers, and enhancers, formulated as a 2x concentrated premix for quantitative or semi-quantitative methylation analysis of DNA templates. |
| Intended Use | Quantitative or semi-quantitative DNA methylation analysis of bisulfite-converted DNA templates by probe-based qPCR; suitable for research and molecular diagnostic assay development. |
| Principle / Technology | Methylation probe (MethyProbe) analysis: after bisulfite treatment of the DNA template, methylated CpG sequences remain as CpG, while unmethylated CpG sequences are converted to UpG. Primers and probes are designed against the CpG and UpG regions for qPCR detection, and methylation levels are determined by comparing Ct values; alternatively, primers/probes targeting only the methylated CpG fragment are used for semi-quantitative analysis. The master mix tolerates bisulfite-converted DNA templates (bisulfite-tolerant and uracil-tolerant) and supports methylation-specific amplification with a high success rate. |
| Detection Method | Real-time fluorescence quantitative PCR (probe-based) |
| Sample Type | Bisulfite-converted DNA templates |
| Performance Range / Specifications | Dedicated methylation qPCR reagent: suitable for methylation qPCR of bisulfite-converted DNA templates; bisulfite- and uracil-tolerant, with a high assay success rate High sensitivity: detects 0.1% methylation of the NDRG4, Septin9, and SDC2 genes in a background of 10 ng human genomic DNA Strong stability: performance stable after 2 weeks at 37 °C and after 10 freeze-thaw cycles |
| Sensitivity / LOD | 0.1% methylation of NDRG4, Septin9, and SDC2 in a 10 ng human genomic DNA background |
| Specificity | Supports methylation-specific amplification targeting methylated CpG fragments; specially processed Taq DNA polymerase and enhancers ensure specific amplification of bisulfite-converted templates. |
| Reaction Conditions / Protocol | Standard probe-based qPCR using bisulfite-converted DNA as template; compare Ct values of CpG- versus UpG-targeting assays to determine methylation level, or use methylation-specific primers/probes for semi-quantitative analysis. |
| Components / Formulation | 2x premix containing: Specially processed Taq DNA polymerase Mg2+ dNTPs PCR stabilizers and enhancers |
| Storage Conditions | Store at -25 °C to -15 °C. |
| Shelf Life | 1 year |
| Package Specifications | 100 T; 1000 T; 10000 T |
| Product Form | Liquid 2x master mix |
| Concentration | 2x (supplied); 1x final working concentration |
| Expiration Date / Stability | Valid for 1 year at -25 °C to -15 °C. Performance unaffected after 14 days at 37 °C or 4 °C (accelerated stability testing) and after 10 freeze-thaw cycles. |
| Application Notes / Precautions | Designed for bisulfite-converted DNA templates; tolerant of bisulfite treatment and uracil residues. Design primers/probes according to methylation status (CpG vs. UpG). Use a defined amount of background genomic DNA when assessing low methylation percentages. |
| Methylation Sensitivity | Detects 0.1% methylation (NDRG4, Septin9, SDC2) in a 10 ng human genomic DNA background. |
For research use only, not for clinical use.
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