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| Product Name | CoAmp Multiplex One-Step RT-qPCR Probe Kit (UDG Plus) |
| Catalog No. | RESK-0129 |
| Description | The CoAmp Multiplex One-Step RT-qPCR Probe Kit (UDG Plus) is a multiplex one-step real-time PCR master mix purpose-built for the efficient, simultaneous detection of DNA and RNA targets within a single reaction. By combining a high-performance heat-resistant reverse transcriptase with a hot-start DNA polymerase in a dedicated reaction buffer system, the kit delivers excellent amplification efficiency and balanced detection performance for both DNA and RNA templates, effectively overcoming the amplification bias commonly seen in conventional reagents during DNA/RNA co-detection. The core components include a deeply optimized 5x MP Buffer and a premixed enzyme system. The buffer already contains sufficient Mg2+, dNTPs and other amplification essentials, together with specific amplification enhancers and multiplex reaction stabilizers, supporting robust multiplex detection of up to four targets while maintaining high amplification efficiency. The built-in dUTP/UDG system efficiently degrades carryover PCR products, fundamentally preventing amplicon contamination and ensuring reliable, reproducible results. |
| Intended Use | For research use in molecular diagnostic assay development: simultaneous (one-tube) qualitative/quantitative detection of DNA and RNA pathogen targets, e.g., co-detection of DNA viruses (such as African swine fever virus) and RNA viruses (such as PRRSV) in mixed-infection scenarios. |
| Principle / Technology | One-step RT-qPCR: reverse transcription and quantitative PCR are performed in the same tube, driven by a heat-resistant reverse transcriptase and a hot-start DNA polymerase in a buffer system optimized for balanced DNA/RNA co-amplification without target bias. Carryover contamination control: the reaction contains dUTP and uracil-DNA glycosylase (UDG); UDG degrades dU-containing carryover PCR products before amplification, preventing false positives from amplicon contamination. |
| Detection Method | Probe-based (fluorogenic probe) real-time quantitative PCR; supports multiplex detection of up to 4 targets in one reaction. |
| Sample Type | Extracted DNA and/or RNA templates; pathogen nucleic acids (validated with plasmid and pseudovirus templates, e.g., HPIV, ASFV, PRRSV). |
| Performance Range / Specifications | One tube, dual detection: DNA + RNA pathogens amplified simultaneously in a single well, saving 50% of reaction wells and doubling throughput. One sample, fast answers: DNA and RNA pathogen results obtained from a single reaction in complex mixed-infection scenarios. One reagent, multiple projects: compatible with both DNA- and RNA-based assays, simplifying production and lowering cost for large-scale manufacturing. Strong performance and stability: sensitivity down to 250 copies/mL; performance unchanged after 14-day accelerated storage at 37 °C; supports fast cycling programs; compatible with major real-time qPCR instruments. Validated co-amplification: amplification efficiency above 90% for both DNA (ASFV) and RNA (PRRSV) targets detected in the same well, with no amplification preference. |
| Sensitivity / LOD | 250 copies/mL. |
| Specificity | Balanced amplification of DNA and RNA templates with no amplification bias; both targets show >90% amplification efficiency in one-well co-detection. |
| Reaction Conditions / Protocol | One-step RT-qPCR protocol; compatible with fast cycling programs on mainstream real-time qPCR instruments. |
| Components / Formulation | 5x MP Buffer: contains Mg2+, dNTPs, specific amplification enhancers and multiplex reaction stabilizers. Premixed enzyme system: heat-resistant reverse transcriptase and hot-start DNA polymerase. dUTP and uracil-DNA glycosylase (UDG) for carryover contamination control. |
| Storage Conditions | Store at -25 to -15 °C. |
| Shelf Life | 12 months. |
| Package Specifications | 50 T; 100 T; 1,000 T; 10,000 T |
| Product Form | Liquid master mix (5x buffer plus premixed enzymes). |
| Quality Control | Stability verified by accelerated testing: after 7 and 14 days at 37 °C, detection rate, Ct values and peak shape for serially diluted plasmid and pseudovirus templates are indistinguishable from reagent stored at -20 °C. |
| Expiration Date / Stability | Valid for 12 months when stored at -25 to -15 °C; performance (detection rate, Ct value, peak shape) unchanged after 14-day accelerated storage at 37 °C. |
| Compatibility | Compatible with major brands of real-time quantitative PCR instruments; supports fast programs. |
| Recommended Buffer System | Supplied 5x MP Buffer (contains Mg2+, dNTPs, amplification enhancers and multiplex stabilizers); no additional buffer required. |
| Application Notes / Precautions | Supports multiplex detection of up to four targets; suitable for both DNA- and RNA-based projects with a single reagent. The dUTP/UDG anti-contamination system is built in; include a UDG digestion step as required by the protocol. |
For research use only, not for clinical use.
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