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| Product Name | Universal Multiplex One-Step RT-qPCR Probe Kit (UDG Plus) |
| Catalog No. | RESK-0087 |
| Description | This kit is designed for multiplex quantitative PCR using RNA as the template. Reverse transcription and quantitative PCR are performed in the same reaction tube, simplifying the workflow. Uracil-DNA glycosylase (UDG) is added to effectively eliminate the risk of aerosol-borne carryover contamination. First-strand cDNA is efficiently synthesized by a thermostable reverse transcriptase, followed by quantitative amplification with a hot-start Taq DNA polymerase. The kit includes an optimized MP Buffer and an Enzymes Mix; the buffer already contains Mg2+ and dNTPs and is supplemented with factors that suppress non-specific PCR amplification and enhance multiplex qPCR efficiency, enabling up to four-plex reactions while maintaining primer amplification efficiency. |
| Intended Use | For one-step, probe-based multiplex quantification of RNA targets where carryover contamination control is required. Suitable as a raw material for molecular diagnostic assay development and research use. |
| Principle / Technology | One-step RT-qPCR: a thermostable reverse transcriptase synthesizes first-strand cDNA, and an antibody-based hot-start Taq DNA polymerase performs probe-based quantitative amplification in the same tube. A dUTP/heat-labile UDG anti-contamination system degrades uracil-containing carryover amplicons to reduce false positives. |
| Detection Method | Fluorescence-based one-step real-time RT-qPCR (hydrolysis probe detection). |
| Sample Type | RNA templates, including crude or inhibitor-containing samples. Tolerates whole blood at up to 5% of the reaction volume and common residual substances from nucleic acid extraction reagents. |
| Performance Range / Specifications | Up to 4-plex amplification in a single reaction Strong inhibitor tolerance - compatible with 5% blood and common extraction reagent residues dUTP/heat-labile UDG anti-contamination system - efficiently degrades aerosol-borne carryover amplicons and reduces false positives Excellent stability - consistent performance over 18 months of real-time monitoring Broad platform compatibility - validated on mainstream real-time PCR instruments |
| Sensitivity / LOD | Reliable detection of 5 copies per reaction. |
| Specificity | The genetically engineered antibody-based hot-start Taq DNA polymerase and buffer additives suppress non-specific amplification, while the dUTP/UDG system eliminates carryover amplicons to reduce false positives. |
| Reaction Conditions / Protocol | One-tube protocol: a UDG digestion step degrades uracil-containing carryover products, followed by reverse transcription and qPCR amplification in the same reaction tube without intermediate handling. Add RNA template, primers and probes to the premixed buffer and Enzymes Mix, then run the programmed thermal profile. |
| Components / Formulation | Optimized MP Buffer (containing Mg2+, dNTPs with dUTP, and factors that suppress non-specific amplification and enhance multiplex amplification efficiency) Enzymes Mix (thermostable reverse transcriptase, antibody-based hot-start Taq DNA polymerase, and heat-labile uracil-DNA glycosylase) |
| Storage Conditions | Store at -25 to -15°C. |
| Shelf Life | 18 months when stored at -25 to -15°C. |
| Package Specifications | 100 T 1000 T 10000 T |
| Product Form | Liquid kit components (buffer and enzyme mix). |
| Quality Control | Performance is verified by sensitivity testing at 5 copies per reaction and by 18-month real-time stability monitoring. |
| Expiration Date / Stability | Stable for 18 months at -25 to -15°C based on real-time stability monitoring. |
| Compatibility | Compatible with mainstream real-time PCR instruments, across mainstream platforms. |
| Recommended Buffer System | Use the supplied optimized MP Buffer, which contains Mg2+, dNTPs/dUTP and multiplex-enhancing additives; no additional buffer is required. |
| Application Notes / Precautions | The dUTP/heat-labile UDG system requires uracil-containing amplicons to be effective; it does not protect against contamination by natural (thymine-containing) template DNA. The one-tube format minimizes handling and reduces contamination risk; standard precautions against RNase contamination should still be applied. The formulation tolerates common extraction reagent residues and up to 5% blood. |
| Heat Inactivation | The heat-labile UDG is inactivated during the subsequent high-temperature steps of the RT-qPCR program, ensuring it does not degrade newly synthesized amplicons. |
For research use only, not for clinical use.
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