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| Product Name | Plus High-Fidelity DNA Polymerase |
| Catalog No. | IVDRM-0099 |
| Description | This enzyme is a genetically engineered high-fidelity DNA polymerase derived from Pyrococcus furiosus DNA polymerase. It exhibits 5'→3' DNA polymerase activity together with 3'→5' exonuclease (proofreading) activity, delivering a fidelity 83-fold higher than Taq DNA polymerase and 9-fold higher than conventional Pfu DNA polymerase. Two monoclonal antibodies in the enzyme formulation suppress both polymerase and proofreading activities at room temperature, enabling convenient, highly specific hot-start PCR with improved detection rates and cleaner target bands. An added elongation factor confers long-fragment amplification capability, allowing targets of up to 13 kb to be amplified. Supplied with an optimized buffer, the enzyme handles complex templates with ease and generates blunt-ended PCR products suitable for downstream cloning. |
| Intended Use | High-fidelity PCR amplification for applications requiring minimal sequence errors, including gene cloning, site-directed mutagenesis, long-fragment amplification, and amplification of complex templates. |
| Principle / Technology | Engineered from Pyrococcus furiosus DNA polymerase; combines 5'→3' DNA polymerase activity with 3'→5' exonuclease proofreading activity to correct misincorporated bases. Hot-start performance is achieved through two monoclonal antibodies that inhibit both activities at ambient temperature, and an elongation factor supports amplification of long targets. |
| Sample Type | Genomic DNA, plasmid DNA, viral DNA, and cDNA templates, including complex and GC-varied templates (GC range 30%–60%). |
| Performance Range / Specifications | Fidelity: 83× Taq DNA polymerase and 9× conventional Pfu DNA polymerase Extension rate: 30 sec/kb; as fast as 10 sec/kb Amplification length: up to 13 kb (λ DNA); ≤10 kb for human gDNA/cDNA Sensitivity: amplifies templates as low as 1 pg Annealing temperature: 68°C GC range: 30%–60% Product ends: blunt ends Stability: no performance loss after 7 days at room temperature or 50 freeze-thaw cycles; room-temperature thawing and pipetting supported |
| Specificity | Antibody-based hot-start mechanism greatly improves amplification specificity and detection rate, yielding a single target band. |
| Reaction Conditions / Protocol | Recommended annealing temperature: 68°C. Extension rate: 30 sec/kb (as fast as 10 sec/kb). 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 | High-Fidelity DNA Polymerase (1 U/μL) 2× PCR Buffer (contains Mg2+ and dNTPs) 6× DNA Loading Buffer |
| Storage Conditions | Store at -20°C. |
| Shelf Life | 1 year. |
| Package Specifications | 10 U 100 U 500 U 1000 U 500 U-EN |
| Product Form | Standalone enzyme |
| Quality Control | Stable performance after 7 days at room temperature and after 50 repeated freeze-thaw cycles. |
| Concentration | 1 U/μL |
| Source / Origin | Genetically engineered enzyme derived from Pyrococcus furiosus DNA polymerase. |
| Shipping Conditions | Shipped with ice packs. |
| Expiration Date / Stability | Stable for 7 days at room temperature and through 50 freeze-thaw cycles without loss of performance; valid for 1 year when stored at -20°C. |
| Compatibility | Suitable for amplification of complex templates; produces blunt-ended products compatible with downstream cloning workflows. |
| Recommended Buffer System | Supplied optimized 2× PCR buffer containing Mg2+ and dNTPs. |
| 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. |
For research use only, not for clinical use.
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