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| Product Name | Universal Multiplex One-Step RT-qPCR Probe Kit (Inhibitor-Tolerant) |
| Catalog No. | RESK-0086 |
| 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 workflow and reducing contamination risk. 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. Suitable as a raw material for developing RT-qPCR assays (e.g., respiratory pathogen and hand-foot-and-mouth disease detection) and for 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. |
| 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 Excellent stability - consistent performance over 18 months of real-time monitoring; stable after 7 days at 37°C even for medium- and low-concentration samples 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, together with buffer factors that suppress non-specific amplification, delivers high amplification specificity. |
| Reaction Conditions / Protocol | One-tube protocol: reverse transcription and qPCR are performed sequentially in the same reaction tube without intermediate handling. Add RNA template, primers and probes to the premixed buffer and Enzymes Mix, then run reverse transcription followed by standard qPCR thermal cycling. |
| Components / Formulation | Optimized MP Buffer (containing Mg2+, dNTPs, and factors that suppress non-specific amplification and enhance multiplex amplification efficiency) Enzymes Mix (thermostable reverse transcriptase and antibody-based 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 | 100 T 500 T 1000 T 10000 T |
| Product Form | Liquid kit components (buffer and enzyme mix). |
| Quality Control | Sensitivity and stability are verified by detection of viral target genes (e.g., ORF and N genes) at 5 copies per reaction with 100% detection across 20 replicates. Stability testing shows reliable amplification of medium- and low-concentration samples (500, 50, and 5 copies per reaction) after 7 days at 37°C. |
| Expiration Date / Stability | Stable for 18 months at -25 to -15°C based on real-time stability monitoring; performance is maintained after 7 days at 37°C. |
| Compatibility | Compatible with mainstream real-time PCR instruments, across mainstream platforms. |
| Recommended Buffer System | Use the supplied optimized MP Buffer, which contains Mg2+, dNTPs and multiplex-enhancing additives; no additional buffer is required. |
| Application Notes / Precautions | Use high-quality RNA templates where possible; the formulation tolerates common extraction reagent residues and up to 5% blood. The one-tube format minimizes handling and reduces contamination risk; standard precautions against RNase contamination should still be applied. |
| Batch-to-Batch Consistency | Verified across production batches: two independent batches showed 100% detection (20/20 replicates) of viral ORF and N gene targets at 5 copies per reaction, with no significant differences in amplification curve profile or fluorescence intensity between batches. |
For research use only, not for clinical use.
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