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| Product Name | qPCR SYBR Green Master Mix (No ROX) |
| Catalog No. | RESK-0005 |
| Description | This 2× real-time quantitative PCR master mix contains hot-start DNA polymerase, SYBR Green I, dNTPs, and Mg2+. Only template and primers need to be added before running real-time quantitative PCR, greatly simplifying the workflow and reducing the risk of contamination. The DNA polymerase ligand adjusts polymerase activity in real time as the temperature changes. The formulation also contains factors that effectively suppress non-specific PCR amplification and enhance amplification efficiency, providing excellent linearity over a wide quantification range. |
| Intended Use | Dye-based real-time quantitative PCR (qPCR) for gene expression analysis and nucleic acid quantification on No ROX instruments. |
| Principle / Technology | Hot-start DNA polymerase whose activity is regulated in real time by a temperature-responsive ligand, combined with SYBR Green I dye-based fluorescence detection. |
| Detection Method | Fluorescent quantitative PCR (SYBR Green dye-based) |
| Sample Type | cDNA and DNA templates for real-time quantitative PCR. |
| Performance Range / Specifications | Ready-to-use format simplifies handling and shortens the workflow. Excellent linearity across a wide quantification range. |
| Specificity | Temperature-responsive polymerase ligand and formulation additives effectively suppress non-specific PCR amplification. |
| Reaction Conditions / Protocol | A two-step cycling protocol is commonly recommended. To improve amplification specificity, use a two-step protocol or increase the annealing temperature; when amplification efficiency is low and CT values are high, switch to a three-step protocol or extend the extension time. Avoid excessively long pre-denaturation. Recommended amplicon length: 80–300 bp. |
| Components / Formulation | Hot-start DNA Polymerase SYBR Green I dNTPs Mg2+ |
| Storage Conditions | Store at -20°C, protected from light. Avoid repeated freeze-thaw cycles. The product contains the fluorescent dye SYBR Green I; protect from strong light during storage and reaction setup. |
| Shelf Life | 18 months when stored at -20°C, protected from light. |
| Package Specifications | 1 mL 5× 1 mL 50× 1 mL 100× 1 mL |
| Product Form | Premixed solution (master mix) |
| Concentration | 2× |
| Shipping Conditions | Shipped with ice packs. |
| Expiration Date / Stability | Stable for 18 months at -20°C protected from light; avoid repeated freeze-thaw cycles. |
| Compatibility | Compatible with qPCR instruments that require No ROX. ROX reference dye can be added separately if the mix is later used on instruments that require ROX. Versions with different premixed ROX concentrations should be selected according to the instrument model. |
| Application Notes / Precautions | Not recommended for non-coding RNAs such as microRNA or lncRNA, or for circular RNA. Avoid excessively long pre-denaturation (e.g., extending 5 min to 10 min), as prolonged pre-denaturation may reduce enzyme activity and lower PCR product yield. A two-step protocol is commonly used; for higher specificity, use a two-step protocol or raise the annealing temperature; for low efficiency/high CT, switch to a three-step protocol or extend the extension time. Valid results require: amplification efficiency of 90–110% (slope -3 to -3.5, R2 > 0.98), S-shaped amplification curves with CT values between 15 and 35 (negative control CT > 35 or absent), and a single melt-curve peak. Minor Tm differences (within 1°C) between replicate wells of the same amplicon are acceptable. CT values are generally inversely correlated with starting template concentration; if inhibitors or impurities are present, diluting the template can lower the CT value. The No ROX master mix can be supplemented with ROX separately for use on instruments that require it. |
For research use only, not for clinical use.
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