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| Product Name | 2x Multiplex ARMS qPCR Mix |
| Catalog No. | RESK-0102 |
| Description | 2x Multiplex ARMS qPCR Mix is a ready-to-use fluorescent quantitative PCR premix with strong amplification-blocking capability, enabling efficient gene genotyping based on ARMS-PCR. The buffer and Taq DNA polymerase are specifically optimized, and the mix contains all ions and dNTPs required for amplification, with no additional components needed. The mix offers high selectivity for detecting a wide range of gene mutations, high sensitivity (mutant primers can detect as little as 0.05% mutation in 10 ng/uL DNA), and straightforward result interpretation without manual ΔCt calculation. |
| Intended Use | Multiplex ARMS-PCR-based qualitative detection and genotyping of gene mutations (e.g., SNPs and hotspot mutations such as KRAS G13D and EGFR L858R) by real-time fluorescence qPCR. |
| Principle / Technology | Amplification Refractory Mutation System (ARMS) PCR: allele-specific primers are designed so that efficient extension occurs only when the primer 3' terminus is perfectly complementary to the mutant (or wild-type) allele. The mix exhibits strong blocking (refractory) capability against mismatched templates, so wild-type templates show no amplification peak or a Ct difference of ≥6-8 versus mutant templates, allowing clear discrimination between wild-type and mutant genotypes. |
| Detection Method | Real-time fluorescence quantitative PCR (qPCR) |
| Sample Type | Genomic DNA; validated with 10 ng/uL DNA inputs (5 uL template per reaction in performance testing) |
| Performance Range / Specifications | High selectivity: detects multiple types of gene mutations. High sensitivity: mutant primers detect as little as 0.05% mutation in 10 ng/uL DNA. Easy interpretation: intuitive, objective results without manual ΔCt calculation. Excellent stability: amplification and blocking performance unaffected after 10 freeze-thaw cycles or 7 days at 37°C. |
| Sensitivity / LOD | Detects 0.05% mutant allele in a 10 ng/uL wild-type DNA background (validated with a 50% self-mixed standard prepared from 10 ng/uL wild-type DNA and 3x10^3 copies of KRAS G13D (38G>A) mutant plasmid). |
| Specificity | In validation testing (5 uL template input), mutant primers amplified 10 ng/uL wild-type template and an equal-copy mutant plasmid, and wild-type primers amplified both templates. Wild-type templates showed no amplification peak or a Ct difference of ≥6-8 versus mutant templates, successfully distinguishing wild-type from mutant genotypes. |
| Reaction Conditions / Protocol | Use as a 2x premix; add primers, template DNA, and water to 1x final concentration. No additional ions or dNTPs are required. Interpret results directly from amplification curves without manual ΔCt calculation. |
| Components / Formulation | 2x premix containing optimized reaction buffer, optimized Taq DNA polymerase, and all ions and dNTPs required for amplification. |
| Storage Conditions | Store at -25 to -15°C. |
| Shelf Life | 1 year when stored at -25 to -15°C. |
| Package Specifications | 100 T 1000 T 10000 T |
| Product Form | 2x concentrated liquid premix |
| Quality Control | Functionally validated for ARMS-PCR genotyping performance, including blocking efficiency, 0.05% mutation detection rate, freeze-thaw stability (10 cycles), and accelerated stability (7 days at 37°C, L858R site). |
| Concentration | 2x |
| Expiration Date / Stability | Valid for 1 year when stored at -25 to -15°C. Amplification and blocking performance remain unaffected after 10 repeated freeze-thaw cycles or storage at 37°C for 7 days. |
| Compatibility | Compatible with standard real-time fluorescence qPCR instruments; suitable for multiplex mutation genotyping assay development. |
| Recommended Buffer System | Built-in optimized buffer system (included in the 2x premix); no additional buffer required. |
| Application Notes / Precautions | The mix contains all ions and dNTPs required for amplification; do not add extra components. Results are read directly from amplification curves; a Ct difference of ≥6-8 between wild-type and mutant templates indicates successful discrimination. |
For research use only, not for clinical use.
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