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| Product Name | 2× Plus High-Fidelity PCR Master Mix (With Dye) |
| Catalog No. | RESK-0032 |
| Description | This product is a ready-to-use 2× premixed solution containing a plus-grade high-fidelity DNA polymerase, dNTPs, and an optimized buffer system. Two monoclonal antibodies in the premix suppress both the polymerase activity and the 3'→5' exonuclease activity at room temperature, enabling convenient, highly specific hot-start PCR. An added elongation factor provides long-fragment amplification capability for targets up to 13 kb. The enzyme exhibits 5'→3' DNA polymerase activity and 3'→5' exonuclease (proofreading) activity, with a fidelity 83-fold higher than Taq DNA polymerase and 9-fold higher than conventional Pfu DNA polymerase. It is suitable for complex templates and produces blunt-ended products. The master mix is fast, convenient, highly sensitive, specific, and stable: only primers and template need to be added, and a two-step PCR program can be used, streamlining the workflow and saving time. An electrophoresis tracking dye is included, so PCR products can be loaded directly onto gels. A specific protective agent keeps the premix active after repeated freeze-thaw cycles. |
| Intended Use | High-fidelity PCR amplification for gene cloning, site-directed mutagenesis, long-fragment amplification, and accurate amplification of complex templates, with direct gel loading of products. |
| Principle / Technology | Based on a high-fidelity DNA polymerase whose 3'→5' exonuclease proofreading activity corrects misincorporated bases during 5'→3' synthesis. Hot-start performance is achieved through two monoclonal antibodies that inhibit both polymerase and proofreading activities at ambient temperature; an elongation factor supports fast, long-fragment amplification. |
| Sample Type | DNA templates including genomic DNA, plasmid DNA, and cDNA, including complex templates with GC contents of 30%–60%. |
| Performance Range / Specifications | Fidelity: 83× Taq DNA polymerase and 9× conventional Pfu DNA polymerase Extension rate: 30 sec/kb efficiently amplifies most fragments; extreme amplification speed of 10 sec/kb Amplification length: targets up to 13 kb; human gDNA/cDNA ≤10 kb; λ DNA ≤13 kb Sensitivity: amplifies templates as low as 1 pg Annealing temperature: 68°C GC range: 30%–60% Product ends: blunt ends Electrophoresis tracking dye: included (direct gel loading) Stability: no performance loss after 7 days at 37°C or 50 freeze-thaw cycles |
| Specificity | Antibody-based hot-start DNA polymerase increases reaction specificity, yielding a single target band. |
| Reaction Conditions / Protocol | 2× ready-to-use format: add only primers and template; room-temperature setup is supported. A two-step PCR program can be used. Recommended annealing temperature: 68°C; extension rate 30 sec/kb (extreme 10 sec/kb). Products contain tracking dye and can be loaded directly for electrophoresis. Recommended template input: genomic DNA 50–200 ng (for 1–10 kb targets); plasmid/viral DNA 10 pg–20 ng; cDNA 1–2.5 μL (no more than 1/10 of the total reaction volume). |
| Components / Formulation | 2× ready-to-use premixed solution containing plus-grade high-fidelity DNA polymerase, dNTPs, and an optimized buffer system; contains two hot-start monoclonal antibodies, an elongation factor, an electrophoresis tracking dye, and a specific protective agent. |
| Storage Conditions | Store at -25°C to -15°C. |
| Shelf Life | 1 year. |
| Package Specifications | 1 mL 100 μL 5× 1 mL |
| Product Form | Master mix (2× premix, with dye) |
| Quality Control | No loss of performance after storage at 37°C for 7 days or after 50 repeated freeze-thaw cycles. |
| Concentration | 2× |
| Expiration Date / Stability | Stable for 7 days at 37°C and through 50 freeze-thaw cycles without loss of performance; valid for 1 year when stored at -25°C to -15°C. |
| Compatibility | Suitable for amplification of complex templates; PCR products contain tracking dye and can be loaded directly for gel electrophoresis. |
| Recommended Buffer System | Built-in optimized buffer system supplied at 2× concentration. |
| Application Notes / Precautions | Recommended template input: genomic DNA 50–200 ng (1–10 kb targets); plasmid/viral DNA 10 pg–20 ng; cDNA 1–2.5 μL, not exceeding 1/10 of the total reaction volume. The formulation contains a specific protective agent and retains stable activity after repeated freeze-thaw cycles; however, aliquoting is recommended to minimize freeze-thaw events. The ready-to-use premix supports room-temperature pipetting, ensuring better reproducibility and accuracy. |
| Batch-to-Batch Consistency | The ready-to-use premixed format supports room-temperature setup and ensures better reproducibility and accuracy between reactions. |
For research use only, not for clinical use.
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