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| Product Name | Lyo-Ready I Multiplex One-Step RT-qPCR Kit (UDG Plus) |
| Catalog No. | RESK-0112 |
| Description | This kit performs multiplex quantitative PCR using RNA as the template. Reverse transcription and quantitative PCR are carried out in the same reaction tube, simplifying the workflow and reducing contamination risk. The unique buffer and enzyme design allows formulation of lyophilized one-step RT-qPCR reaction systems and product development. A thermostable reverse transcriptase efficiently synthesizes first-strand cDNA, and an antibody-based hot-start Taq DNA polymerase performs quantitative amplification. The kit includes an optimized lyophilization buffer and an enzyme mix containing factors that suppress non-specific PCR amplification and improve multiplex qPCR efficiency, enabling multiplex real-time amplification while maintaining primer amplification efficiency. A lyoprotectant is included, so the reagent can be lyophilized directly. |
| Intended Use | One-step multiplex probe-based RT-qPCR using RNA templates; designed for development and production of lyophilized RT-qPCR assays, and can also be used directly as a liquid reagent. |
| Principle / Technology | Thermostable reverse transcriptase synthesizes first-strand cDNA, followed by quantitative amplification with a genetically engineered antibody-based hot-start Taq DNA polymerase and hydrolysis probes in a single tube. The formulation contains a lyoprotectant enabling direct freeze-drying; enzyme activity remains fully intact after lyophilization, with performance equivalent to the liquid reagent. A dUTP/UDG system prevents aerosol carryover contamination. |
| Detection Method | Real-time fluorescence quantitative PCR (probe-based, multiplex) |
| Sample Type | RNA (e.g., transcribed RNA templates; extracted RNA) |
| Performance Range / Specifications | Enzyme activity fully preserved after lyophilization; performance equivalent to the liquid reagent High sensitivity with genetically engineered antibody hot-start Taq: 5 copies/T stably detected Lyophilized beads containing primers and probes: performance unaffected after 1 month of accelerated heating at 50 °C Performance unaffected under fast protocols: results in 30 min dUTP/UDG anti-contamination system effectively prevents aerosol contamination Contains lyoprotectant: can be lyophilized directly or used as a liquid reagent |
| Sensitivity / LOD | 5 copies/T stably detected; low-concentration detection at 1 copy/uL with 100% detection rate (20 replicates) |
| Specificity | Contains factors that suppress non-specific PCR amplification; hot-start Taq polymerase improves amplification specificity. |
| Reaction Conditions / Protocol | Amplification program: 50 °C 2 min; 95 °C 30 sec; 40 cycles (95 °C 5 sec; 60 °C 10 sec) Liquid system: 25 uL total volume with 5 uL template Lyophilized system: 20 uL reaction lyophilized (including primers/probes); reconstitute 1 bead with 20 uL nuclease-free water and add 5 uL template |
| Components / Formulation | Optimized lyophilization buffer Enzymes Mix (thermostable reverse transcriptase + genetically engineered antibody-based hot-start Taq DNA polymerase) Lyoprotectant (enables direct lyophilization) dUTP/UDG anti-contamination system |
| Storage Conditions | Store at -25 °C to -15 °C. |
| Shelf Life | 1 year |
| Package Specifications | 100 T; 1000 T; 10000 T |
| Product Form | Liquid master mix (lyophilization-ready; can be processed into lyophilized beads) |
| Expiration Date / Stability | Valid for 1 year at -25 °C to -15 °C. Lyophilized beads containing primers and probes show no change in amplification curve shape, Ct values, or low-concentration detection rate (1 copy/uL, 100% in 20 replicates) after 1 month at 50 °C. Post-lyophilization reconstituted reagent performance is stable and equivalent to the liquid reagent. |
| Compatibility | Validated on common real-time PCR instruments ; compatible with fast cycling programs (results in 30 min). |
| Application Notes / Precautions | For lyophilized formats, reconstitute each bead with nuclease-free water before adding template. Use the dUTP/UDG step to control carryover contamination. Follow recommended freeze-drying process and conditions for bead production. |
| Batch-to-Batch Consistency | Lyophilized beads from accelerated stability testing show no difference versus freshly produced beads from the same batch. |
For research use only, not for clinical use.
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