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| Product Name | Antibody Hot-Start qPCR SYBR Green Master Mix (High ROX) |
| Catalog No. | RESK-0014 |
| Description | This product is a 2× premixed solution for real-time quantitative PCR amplification. It employs an antibody-based hot-start DNA polymerase: during the 95°C pre-denaturation step (30 sec), the anti-Taq antibody is inactivated, releasing Taq DNA polymerase activity. Antibody hot-start Taq effectively suppresses amplification caused by non-specific primer annealing. The formulation is optimized with factors that inhibit non-specific PCR amplification and boost amplification efficiency, making it well suited for low-concentration templates and enabling robust standard curves across a wide quantitative range. The mix contains hot-start Taq DNA polymerase, anti-Taq antibody, SYBR Green I, dNTPs, Mg2+, and High ROX reference dye. Only template and primers need to be added to run real-time fluorescent quantitative PCR, greatly simplifying the workflow and reducing the risk of contamination. Developed as a high-sensitivity formulation, it is optimized for low-abundance samples. |
| Intended Use | For dye-based (SYBR Green I) real-time fluorescent quantitative PCR. Suitable for amplification of low-concentration templates and for generating reliable standard curves over a wide quantitative range. This high-sensitivity version is developed and optimized for low-abundance samples. |
| Principle / Technology | Antibody-mediated hot-start technology: the anti-Taq antibody blocks polymerase activity during reaction setup; heating at 95°C for 30 sec during pre-denaturation irreversibly inactivates the antibody and releases Taq DNA polymerase activity. This effectively suppresses amplification caused by non-specific primer annealing. The formulation also contains factors that inhibit non-specific PCR amplification and enhance amplification efficiency. |
| Detection Method | Fluorescent quantitative PCR (SYBR Green dye-based) |
| Sample Type | DNA or cDNA templates. A common input is cDNA reverse-transcribed from 500-1000 ng total RNA, diluted 10-fold, with 1 µL used per qPCR reaction. |
| Performance Range / Specifications | High specificity: antibody hot-start suppresses non-specific amplification. High sensitivity: developed and optimized for low-abundance samples. Suitable for amplification of low-concentration templates; reliable standard curves over a wide quantitative range. Recommended amplification length: 80-300 bp. |
| Sensitivity / LOD | High-sensitivity formulation optimized for low-abundance samples; suitable when Ct values consistently exceed 30 with other reagents. |
| Specificity | The antibody hot-start Taq polymerase effectively suppresses amplification resulting from non-specific primer annealing, delivering high reaction specificity. |
| Reaction Conditions / Protocol | Only template and primers need to be added to the 2× premix. Hot-start activation: 95°C for 30 sec (inactivates the anti-Taq antibody). Optimal template input should be determined empirically: dilute the template (generally 5-10 fold) and run a template gradient, selecting the input that gives a Ct value of 20-30. Recommended cycle number: generally 40; up to 45 for low-abundance targets. Recommended amplification length: 80-300 bp. For detailed cycling parameters, refer to the product manual. |
| Components / Formulation | 2× qPCR SYBR Green Master Mix (High ROX), containing: Hot-start Taq DNA polymerase Anti-Taq antibody SYBR Green I dNTPs Mg2+ High ROX reference dye |
| Storage Conditions | Store at -20°C protected from light. Avoid repeated freeze-thaw cycles. The product contains the fluorescent dye SYBR Green I; avoid exposure to strong light during storage and reaction setup. |
| Shelf Life | 18 months. |
| Package Specifications | 1 mL; 5×1 mL; 50×1 mL; 100×1 mL |
| Product Form | Premix (2× master mix) |
| Concentration | 2× |
| Shipping Conditions | Shipped with ice packs. |
| Expiration Date / Stability | Valid for 18 months when stored at -20°C protected from light. Avoid repeated freeze-thaw cycles and exposure to strong light (the mix contains SYBR Green I). |
| Compatibility | Contains High ROX reference dye; compatible with real-time qPCR instruments that require High ROX. |
| Application Notes / Precautions | This high-sensitivity formulation is developed and optimized for low-abundance samples; it is a suitable choice when Ct values consistently exceed 30 in relative quantification experiments. Determine optimal template input empirically: dilute the template (generally 5-10 fold) and run a gradient, choosing the input that yields a Ct of 20-30. Avoid repeated freeze-thaw cycles, and protect from strong light during storage and reaction setup because the mix contains SYBR Green I. |
For research use only, not for clinical use.
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