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Product Overview
This product is a special reagent for Real Time One Step RT-PCR using the SYBR Green method. Using this product for Real Time RT-PCR reaction can continuously perform reverse transcription and PCR amplification in the same reaction tube. It is simple to operate and can effectively prevent contamination. Since this reaction system can monitor the amplified products in real time, the detection sensitivity is greatly improved, and the electrophoresis step after PCR reaction is omitted, which is very suitable for the detection of RNA viruses. This product uses high-efficiency reverse transcriptase and high-specificity hot-start Taq DNA polymerase to perform stable and efficient Real Time One Step RT-PCR reactions. For the fluorescent quantitative PCR instrument that uses ROX as the calibration dye, this product is equipped with a separate ROX dye to correct the fluorescent signal error generated between the wells of the quantitative PCR instrument.
Total RNA input is 1 pg–1 µg per reaction and reactions are run in 20 µL or 50 µL, with amplified products of 80–250 bp and a final primer concentration of 0.2 µM–0.6 µM. The program is reverse transcription at 50°C for 15 min, pre-denaturation at 95°C for 2 min, then 35–40 cycles of 95°C for 10 sec and 60°C for 30 sec, followed by melt curve analysis.
Specifications
| Format | 2× one-step RT-PCR mix with separate RT-PCR Enzyme Mix |
|---|---|
| Detection chemistry | SYBR Green method |
| Reverse transcriptase | High-efficiency third-generation reverse transcriptase |
| Polymerase | High-specificity hot-start Taq DNA polymerase |
| ROX | Separate 50× ROX Dye supplied |
| Template input | Total RNA 1 pg–1 µg |
| Reaction volume | 20 µL or 50 µL |
| Amplified product length | 80–250 bp |
Kit Components
| Component | 20 µL × 100 rxn | 20 µL × 500 rxn |
|---|---|---|
| 2×One Step RT-PCR Mix (SYBR Green) | 1 mL | 1.25 mL × 4 |
| RT-PCR Enzyme Mix | 150 µL | 750 µL |
| 50×ROX Dye | 250 µL | 1.25 mL |
| RNase-free ddH2O | 1 mL | 1.25 mL × 4 |
Applications
- Real-time one-step RT-PCR
- Detection of viral RNA in research samples
- Detection of trace RNA
Key Features
- Add RNA only: reverse transcription and PCR amplification run consecutively in the same PCR tube, keeping the operation simple.
- Real-time detection of amplified products greatly improves detection sensitivity and can effectively prevent contamination.
- The preferred third-generation reverse transcriptase is used in conjunction with the hot-start Taq enzyme to obtain optimal amplification and effectively reduce non-specific amplification.
- The electrophoresis step after the PCR reaction is omitted, which is very suitable for the detection of trace RNA.
Storage & Handling
All components are stored at -20°C with a shelf life of 2 years. Avoid strong light.
Usage Notes
- Do not vortex the reagents.
- Minimize freeze-thaw cycles.
- Use filtered tips.
- Keep reagents on ice.
- Avoid strong light.
Safety & Regulatory
For research use only. Not intended for clinical, diagnostic or therapeutic use. Follow the manufacturer's manual and standard laboratory safety practice.
Customization & Add-ons: Need this molecular biology reagent tailored to your workflow? We can help with bulk or custom pack sizes, alternative formats (spin column, magnetic bead or reagent-only kits; premixed or stand-alone reference dye), matched extraction, cDNA synthesis and qPCR workflows, and sourcing of related enzymes, ladders and consumables not in our catalog. Click Talk to a Scientist to submit a request, email us at support@biohippo.com, or explore our Research Services — our team will follow up with feasibility details and next steps.
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