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| Product Name | 2× Ultra-Rapid II Hot-Start PCR Master Mix |
| Catalog No. | RESK-0046 |
| Description | The 2× Ultra-Rapid II Hot-Start PCR Master Mix is built around an antibody-inactivated, thermostable Taq DNA polymerase, enhanced with a potent elongation factor and an optimized buffer system to deliver exceptionally high amplification efficiency. It is ideally suited for colony PCR of E. coli, Agrobacterium, yeast, and most other microorganisms. For complex templates such as genomic DNA, extension rates reach 1 sec/kb for targets up to 3 kb, 3 sec/kb for 3-6 kb fragments, 5 sec/kb for 6-10 kb fragments, and 10 sec/kb for fragments over 10 kb. Simple templates such as plasmids up to 6 kb can be extended at 1 sec/kb, dramatically shortening total PCR run time. The premixed format contains dNTPs and Mg2+, so only primers and template need to be added. A red tracking dye is included for direct gel loading after amplification, and built-in protectants keep the enzyme fully active even after repeated freeze-thaw cycles. PCR products carry a 3'-dA overhang and can be cloned directly into T vectors. |
| Intended Use | Ultra-rapid PCR, colony PCR (E. coli, Agrobacterium, yeast, and other microorganisms), and gene identification across human, mouse, plant, fungal, and bacterial templates. |
| Principle / Technology | Antibody-mediated hot-start technology: the thermostable Taq DNA polymerase is blocked by an antibody modifier at low temperature and activated upon initial denaturation. A strong elongation factor and an optimized buffer system further boost amplification speed and efficiency. |
| Sample Type | Human, mouse, plant, fungal, and bacterial samples; suitable for colonies, crude bacterial cultures, genomic DNA, plasmid DNA, and lambda DNA. |
| Performance Range / Specifications | Extension speed: 1 sec/kb for complex templates (genomic DNA) up to 3 kb; 3 sec/kb for 3-6 kb; 5 sec/kb for 6-10 kb; 10 sec/kb for fragments over 10 kb; 1 sec/kb for simple templates (plasmids) up to 6 kb. Amplicon length: up to 15 kb (lambda DNA), up to 10 kb for genomic DNA, up to 10 kb for colonies and bacterial cultures. GC compatibility: 30-70% GC content. Detection rate: 100% for 1-5 kb E. coli colonies and 1-10 kb Agrobacterium colonies. Validated on hundreds of fragments across more than ten species. Annealing temperature: 60°C. |
| Sensitivity / LOD | Detection rate of up to 100% across validated targets, including 1-5 kb E. coli colony fragments, 1-10 kb Agrobacterium colony fragments, and 1-10 kb mouse genomic DNA. |
| Specificity | High amplification specificity with detection rates up to 100%; outperforms competing products in yield and detection rate across colony, genomic, lambda DNA, and low-to-high GC templates. |
| Reaction Conditions / Protocol | Annealing temperature: 60°C. Extension time: 1-10 sec/kb depending on template type and amplicon length (e.g., 1 sec/kb for targets up to 3 kb, 10 sec/kb for fragments over 10 kb). Add only primers and template to the 2× mix. |
| Components / Formulation | Antibody-modified thermostable Taq DNA polymerase, dNTPs, Mg2+, elongation factor, optimized buffer system, red tracking dye, and stabilizing protectants. Catalog numbers of individual components are not applicable. |
| Storage Conditions | Store at -25°C to -15°C. |
| Shelf Life | 2 years when stored at -25°C to -15°C. |
| Package Specifications | 1 mL; 5× 1 mL; 50× 1 mL; 100× 1 mL; 10× 5 mL |
| Product Form | Premixed master mix (Mix) |
| Quality Control | Performance verified by amplification of colony, genomic DNA, lambda DNA, and low-to-high GC templates with detection rates up to 100%. |
| Concentration | 2× |
| Source / Origin | Thermostable Taq DNA polymerase modified with a hot-start antibody. |
| Expiration Date / Stability | Stable for 2 years at -25°C to -15°C. Retains full activity after 100 freeze-thaw cycles and after incubation at 37°C for 14 days. |
| Compatibility | Suitable for templates with 30-70% GC content and amplicons up to 15 kb; PCR products carry 3'-dA overhangs compatible with T-vector (TA) cloning. |
| Application Notes / Precautions | Only primers and template need to be added; no additional dNTPs or Mg2+ required. The mix contains a red tracking dye, allowing direct gel loading after PCR. PCR products carry 3'-dA overhangs and can be cloned directly into T vectors. The formulation tolerates repeated freeze-thaw cycles without loss of activity. |
| Batch-to-Batch Consistency | Validated on hundreds of fragments from more than ten species with detection rates up to 100%; stable performance maintained after 100 freeze-thaw cycles and 14 days at 37°C. |
For research use only, not for clinical use.
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