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| Product Name | C58P2 Multiplex One-Step RT-qPCR Probe Kit (UDG Plus) |
| Catalog No. | RESK-0100 |
| Description | The C58P2 Multiplex One-Step RT-qPCR Probe Kit (UDG Plus) is designed for multiplex quantitative PCR using RNA templates. Reverse transcription and quantitative PCR are performed in the same reaction tube, simplifying operation and reducing contamination risk. A thermostable reverse transcriptase efficiently synthesizes first-strand cDNA, and an antibody-based hot-start Taq DNA polymerase drives quantitative amplification. The kit includes an optimized MP buffer and an enzyme mix; the buffer already contains Mg2+ and dNTPs plus factors that suppress non-specific amplification and enhance multiplex amplification efficiency, enabling multiplex amplification while maintaining primer efficiency. A dUTP/UDG system is included to effectively prevent aerosol contamination. |
| Intended Use | Multiplex one-step RT-qPCR detection of RNA targets for molecular diagnostic reagent development and research applications. |
| Principle / Technology | One-step RT-qPCR: reverse transcription and quantitative PCR are performed sequentially in the same reaction tube. A thermostable reverse transcriptase efficiently synthesizes first-strand cDNA, followed by quantitative amplification with an antibody-based hot-start Taq DNA polymerase. The incorporated dUTP/UDG anti-contamination system degrades uracil-bearing carryover amplicons, effectively preventing aerosol contamination and reducing false positives. |
| Detection Method | TaqMan probe-based one-step real-time fluorescence RT-qPCR. |
| Sample Type | RNA templates. |
| Performance Range / Specifications | Excellent amplification sensitivity: detection of low-concentration templates down to 250 copies/mL. Outstanding storage stability: passes 18-month real-time stability testing. dUTP/UDG anti-contamination system efficiently degrades aerosol-borne product carryover, reducing false positives. Supports fast programs: results within 40 min. Supports multiplex (up to 4-plex) amplification. |
| Sensitivity / LOD | 250 copies/mL. In validation with serially diluted pseudovirus (500 and 250 copies/mL, ORF and N gene targets), the detection rate was 100% across 16 replicate wells per concentration. |
| Specificity | The buffer contains factors that effectively suppress non-specific PCR amplification and factors that enhance amplification efficiency in multiplex qPCR, ensuring primer efficiency in up to 4-plex reactions. |
| Reaction Conditions / Protocol | One-step protocol: reverse transcription and qPCR in a single closed tube. Compatible with both conventional and fast cycling programs; with the fast program, detection is completed within 40 min. In 4-plex reactions, amplification curves and Ct values are equivalent between conventional and fast programs. |
| Components / Formulation | Optimized MP buffer (contains Mg2+ and dNTPs, with dUTP/UDG system) Enzyme mix (thermostable reverse transcriptase and hot-start Taq DNA polymerase) |
| Storage Conditions | Store at -25 to -15°C. |
| Shelf Life | 18 months when stored at -25 to -15°C. |
| Package Specifications | 50 T 100 T 1000 T 5000 T 10000 T |
| Product Form | Liquid one-step RT-qPCR kit with separate buffer and enzyme mix components. |
| Quality Control | Sensitivity validation: 100% detection rate at 500 and 250 copies/mL (16 replicates each). Stability validation: batches stored 18 months at -20°C show amplification curve shape and Ct values equivalent to in-date reagents, with 100% detection at 1000 copies/mL (20 replicates). |
| Expiration Date / Stability | Valid for 18 months when stored at -25 to -15°C. Real-time stability verified over 18 months. Performance is unaffected after 30 or 50 repeated freeze-thaw cycles (on dry ice) compared with reagents stored continuously at -20°C. |
| Regulatory / Compliance | For research use and in vitro diagnostic reagent development. Not for direct clinical diagnosis. |
| Compatibility | Compatible with both conventional and fast cycling programs on mainstream real-time PCR instruments; equivalent performance demonstrated in 4-plex systems. |
| Recommended Buffer System | Supplied optimized MP buffer containing Mg2+, dNTPs and the dUTP/UDG system. |
| Application Notes / Precautions | Handle RNA templates under RNase-free conditions. Include the UDG digestion step in the cycling program to ensure carryover control. |
| Batch-to-Batch Consistency | Batches stored for 18 months at -20°C show no significant difference in amplification curve shape or Ct value compared with in-date batches (1000 copies/mL, 20 replicates, 100% detection). |
For research use only, not for clinical use.
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