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| Product Name | III One-Step RT-qPCR SYBR Green Kit |
| Catalog No. | RESK-0067 |
| Description | The III One-Step RT-qPCR SYBR Green Kit is designed for quantitative PCR using RNA templates. Reverse transcription and quantitative PCR proceed in the same reaction tube, simplifying handling and reducing contamination risk. Using gene-specific primers, a thermostable reverse transcriptase efficiently synthesizes first-strand cDNA, which is then amplified by an antibody-based hot-start Taq DNA polymerase. Together with an optimized buffer system, the kit enables specific and sensitive quantification of RNA templates. The buffer additionally contains components that effectively suppress nonspecific PCR amplification and enhancing factors that improve amplification capability, ensuring accurate and reliable quantitative results over a wide dynamic range. The kit is supplied in a convenient format: the 2x SG buffer contains the optimized buffer system and dNTPs, optimized with factors that suppress nonspecific amplification and boost PCR efficiency, and is suited to SYBR Green-based high-specificity detection. The UH III Enzymes blend contains an optimally proportioned mix of reverse transcriptase, RNase inhibitor, and hot-start Taq DNA polymerase. |
| Intended Use | Quantitative PCR with RNA templates; particularly suited to virus detection applications requiring a high detection rate. |
| Principle / Technology | One-tube reverse transcription and qPCR with gene-specific primers; thermostable reverse transcriptase for first-strand cDNA synthesis; antibody-based hot-start Taq DNA polymerase; buffer optimized with nonspecific-amplification inhibitors and amplification-enhancing factors. |
| Detection Method | SYBR Green dye-based fluorescence detection. |
| Sample Type | RNA templates, including viral RNA. |
| Performance Range / Specifications | Detection rate: >90%. Amplification efficiency: >90%. Strong specificity: no nonspecific amplification. High sensitivity: detects >10 pg total RNA; GAPDH reliably detected from as little as 2 pg input. Storage stability: buffer withstands >50 freeze-thaw cycles when stored separately from the enzyme mix. Annealing temperature 60°C; GC range 30%-70%; amplicon length 80-300 bp. |
| Sensitivity / LOD | Detects >10 pg total RNA; GAPDH detectable from as little as 2 pg input. |
| Specificity | No nonspecific amplification; lower Ct values and single amplification products compared with competing products across templates from various sources. |
| Reaction Conditions / Protocol | Reverse transcription and qPCR performed sequentially in the same tube; recommended annealing temperature 60°C; amplicon length 80-300 bp. |
| Components / Formulation | 2x III SG Buffer: 500 μL (50 T) / 4x500 μL (200 T) / 5 mL (500 T) UH III Enzymes: 50 μL (50 T) / 200 μL (200 T) / 500 μL (500 T) 50x High ROX: 50 μL (50 T) / 200 μL (200 T) / 500 μL (500 T) 50x Low ROX: 50 μL (50 T) / 200 μL (200 T) / 500 μL (500 T) RNase-free ddH2O: 500 μL (50 T) / 4x500 μL (200 T) / 5 mL (500 T) |
| Storage Conditions | Store at -25°C to -15°C, protected from light. Avoid repeated freeze-thaw cycles; aliquoting is recommended. |
| Shelf Life | 1 year when stored at -25°C to -15°C, protected from light. |
| Package Specifications | 50 T; 200 T; 500 T |
| Product Form | Mix |
| Quality Control | Detection rate >90% and amplification efficiency >90% verified; sensitivity validated down to 2 pg input for GAPDH. |
| Expiration Date / Stability | 1 year at -25°C to -15°C protected from light; buffer is freeze-thaw stable (>50 cycles) when stored separately from the enzymes. |
| Compatibility | Compatible with the vast majority of real-time PCR instruments; ROX reference dye (50x High ROX and 50x Low ROX) is supplied separately and added as required by the instrument. |
| Recommended Buffer System | 2x optimized SYBR Green buffer containing dNTPs, nonspecific-amplification inhibitors, and amplification-enhancing factors (included). |
| Application Notes / Precautions | Designed for virus detection and other applications demanding a high detection rate. Avoid repeated freeze-thaw cycles; aliquot for storage. The buffer is freeze-thaw stable (>50 cycles) when kept separate from the enzyme mix. |
For research use only, not for clinical use.
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