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| Product Name | Universal TaqMan Multiplex qPCR Master Mix (Inhibitor-Tolerant) |
| Catalog No. | RESK-0084 |
| Description | This product is a 2x premixed reagent that enables up to four-plex fluorescence quantitative PCR in a single reaction well. It contains a genetically engineered, antibody-based hot-start Taq DNA polymerase that greatly improves amplification sensitivity and specificity. The multiplex reaction buffer system has been deeply optimized to increase amplification efficiency and promote efficient amplification of low-concentration templates. It is suitable for genotyping and multiplex gene quantification analysis. |
| Intended Use | For probe-based multiplex real-time quantitative PCR applications, including genotyping and multiplex gene quantification. Suitable as a raw material for molecular diagnostic assay development and research use. |
| Principle / Technology | TaqMan probe-based real-time quantitative PCR using a genetically engineered antibody-mediated hot-start Taq DNA polymerase in an optimized multiplex buffer system. |
| Detection Method | Fluorescence-based real-time quantitative PCR (TaqMan hydrolysis probe detection). |
| Sample Type | DNA templates, including inhibitor-containing crude samples. Tolerates whole blood at up to 5% of the reaction volume and common anticoagulants used in blood collection such as EDTA and sodium citrate. |
| Performance Range / Specifications | Up to 4-plex amplification in a single reaction well Strong tolerance to blood impurities - compatible with samples containing up to 5% blood Excellent storage stability - stable after 2 weeks at 37°C and repeated freeze-thaw cycles Broad platform compatibility - validated on mainstream real-time PCR instruments |
| Sensitivity / LOD | Reliable detection of 3 copies per reaction. |
| Specificity | The genetically engineered antibody-based hot-start Taq DNA polymerase suppresses non-specific amplification and primer-dimer formation, delivering high amplification specificity. |
| Reaction Conditions / Protocol | Recommended template optimization: dilute template DNA (generally 5-10 fold) and run a loading gradient; select the input amount giving a Ct value between 20 and 30. Typical input: cDNA reverse-transcribed from 500-1000 ng total RNA, diluted 10-fold, with 1 uL used per qPCR reaction. Recommended cycle number: 40 cycles (45 cycles for low-abundance targets). For valid results, amplification efficiency should be 90-110% (slope -3.0 to -3.5, R2 > 0.98) with target Ct values between 15 and 35. |
| Components / Formulation | 2x master mix containing a genetically engineered antibody-based hot-start Taq DNA polymerase in an optimized multiplex reaction buffer (with Mg2+ and dNTPs). |
| Storage Conditions | Store at -25 to -15°C. |
| Shelf Life | 2 years when stored at -25 to -15°C. |
| Package Specifications | 1 mL 5 x 1 mL 100 x 1 mL 5 mL 20 mL 100 mL |
| Product Form | Liquid, 2x concentrated premixed master mix. |
| Quality Control | Stability verified by accelerated testing at 37°C for 7 and 14 days and by repeated freeze-thaw testing at -20°C, with no significant changes in Ct values or fluorescence signals. |
| Expiration Date / Stability | Stable for 24 months at -25 to -15°C. Performance is maintained after 2 weeks at 37°C and after repeated freeze-thaw cycles. |
| Compatibility | Compatible with mainstream real-time PCR instruments, across mainstream platforms. Suitable for multiplex probe-based qPCR assays. |
| Recommended Buffer System | The 2x master mix contains a fully optimized multiplex reaction buffer with Mg2+ and dNTPs; no additional buffer is required. |
| Application Notes / Precautions | Optimize template input on first use; target Ct values between 15 and 30 are recommended, as Ct values below 15 may distort baseline setting. ROX reference dye normalizes non-PCR-related fluorescence fluctuations and corrects pipetting or well-to-well variation. Use high-purity RNA (OD260/OD280 = 1.8-2.0, OD260/OD230 > 2.0) to avoid jagged amplification curves, and calibrate instruments regularly (about once per year). If the amplification curve fails to plateau, increase template input or cycle number (up to 45 for low-abundance targets); low amplification efficiency may require reagent replacement. Peaks with Tm below 80°C in melting analysis usually indicate primer-dimers; increase annealing temperature, reduce primer concentration, or redesign primers. |
For research use only, not for clinical use.
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