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| Product Name | KASP Genotyping Master Mix |
| Catalog No. | RESK-0134 |
| Description | This master mix is developed based on kompetitive allele-specific PCR (KASP) technology for rapid genotyping assays. Results are interpreted by endpoint fluorescence scanning, enabling accurate detection of point mutations, insertions, and deletions for fast and reliable genotyping. KASP is a homogeneous, endpoint genotyping technology based on fluorescence signal readout. Unlike conventional TaqMan probe assays, which require a specific fluorescent probe for each SNP locus, this method uses universal probes directed against the tag sequence of the allele-specific upstream primers. The universal probes can be combined with a wide variety of gene-specific primers, eliminating the need to synthesize a dedicated probe for every locus and greatly reducing reagent costs. The format supports low-, medium-, and high-throughput genotyping and is readily adaptable to high-throughput, automated workflows. |
| Intended Use | For rapid, accurate genotyping of SNPs, point mutations, insertions, and deletions in DNA samples; suitable for low-, medium-, and high-throughput genotyping applications. |
| Principle / Technology | KASP is a homogeneous, endpoint genotyping technology that combines allele-specific PCR (AS-PCR) with a fluorescence resonance energy transfer (FRET) signal system. A touchdown PCR program ensures that primers and probes bind as intended: at the initial high annealing temperature, the high-Tm allele-specific primers bind the template specifically and are extended by Taq DNA polymerase, enriching the target template; as the annealing temperature decreases, the universal probes hybridize and generate fluorescence. Endpoint fluorescence is read below 40 °C for SNP calling, ensuring high genotyping accuracy. Two allele-specific forward primers carry distinct tag sequences that are recognized by universal FAM- and HEX-labeled probe cassettes, so the FAM/HEX fluorescence ratio identifies the genotype. |
| Detection Method | Endpoint fluorescence scanning (FAM/HEX dual-channel readout below 40 °C); genotype is assigned from the allele-specific fluorescence signals. |
| Sample Type | DNA templates (e.g., genomic DNA for SNP and indel genotyping) |
| Performance Range / Specifications | Accurately discriminates wild-type, mutant, and mixed templates tested at 100 copies/uL, with clear amplification profiles and reliable genotyping across multiple primer sets. |
| Sensitivity / LOD | Demonstrated detection and correct genotyping of wild-type and mutant templates at 100 copies/uL. |
| Specificity | Allele-specific primers combined with a touchdown PCR program provide highly specific template binding and amplification, ensuring high genotyping accuracy. |
| Reaction Conditions / Protocol | Touchdown PCR program: initial cycles at a high annealing temperature for specific primer binding and amplification, followed by progressively lower annealing temperatures for probe hybridization and fluorescence generation; endpoint fluorescence readout performed below 40 °C. |
| Components / Formulation | Ready-to-use master mix optimized for KASP assays, containing Taq DNA polymerase, universal FAM/HEX fluorescent probe system, reaction buffer, dNTPs, and Mg2+. |
| Storage Conditions | Store protected from light at -20 ± 5 °C. |
| Shelf Life | 12 months; use within the validity period. |
| Package Specifications | 1 mL 5 mL 25 mL |
| Product Form | Liquid master mix |
| Shipping Conditions | Ship with dry ice together with ice packs; total low-temperature transit time should not exceed 5 days. |
| Expiration Date / Stability | Avoid repeated freeze-thaw cycles; stable for 12 months when stored protected from light at -20 ± 5 °C. |
| Regulatory / Compliance | For research use only. |
| Compatibility | Compatible with mainstream real-time PCR instruments and fluorescence plate readers supporting FAM/HEX channels; suitable for automated, high-throughput workflows. |
| Application Notes / Precautions | Protect from light during storage and handling. Avoid repeated freeze-thaw cycles. Use within the shelf life. |
For research use only, not for clinical use.
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