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Product Overview
Hot Start Taq DNA Polymerase is an enzyme solution in which a monoclonal antibody against Taq enzyme and Taq enzyme are mixed in a 1:1 activity unit. Taq enzyme antibody has a very high affinity with Taq enzyme. It can block the activity of Taq enzyme before high temperature denaturation, so it can inhibit primer dimer and non-specific amplification very effectively, greatly improving the accuracy of the PCR reaction. Hot Start Taq DNA Polymerase does not require special high temperature treatment. The pre-denaturation step in conventional PCR reaction conditions can start the activity of Taq enzyme. It is suitable for various Taq enzyme-based hot-start PCR and qPCR reactions. The PCR product has an A at the 3' end, which can be cloned directly with TA vector.
The enzyme is supplied at 5 U/µL together with 10× PCR Buffer (Mg²⁺) and ddH₂O; no separate MgCl₂ is supplied. A 50 µL reaction uses 0.5–1.0 µL of polymerase at a final Mg²⁺ concentration of 2 mM, adjustable between 2 and 5 mM. Cycling is 94°C for 1–5 min, then 25–35 cycles of 94°C for 15 sec, 50–60°C for 30 sec and 72°C for 30 sec, with a final 72°C for 5 min.
Specifications
| Concentration | 5 U/µL |
|---|---|
| Unit definition | Incorporates 10 nmol dNTPs into acid-insoluble material in 30 min at 74°C |
| Hot start mechanism | Monoclonal antibody against Taq enzyme mixed with Taq enzyme in a 1:1 activity unit |
| Activation | No special high temperature treatment; activated by the pre-denaturation step of conventional PCR |
| Purity | Greater than 99% by SDS-PAGE |
| Supplied buffer | 10× PCR Buffer (Mg²⁺) |
| Template input per 50 µL | Human genomic DNA 0.05–0.5 µg; E. coli genomic DNA 10–100 ng; λ DNA 0.5–5 ng; plasmid DNA 0.1–10 ng |
| Product ends | A at the 3' end for direct TA vector cloning |
Kit Components
| Component | 500 U (100 µL) | 2500 U (500 µL) |
|---|---|---|
| Hot Start Taq DNA Polymerase (5 U/µL) | 100 µL | 500 µL |
| 10× PCR Buffer (Mg²⁺) | 1 mL | 5 mL |
| ddH₂O | 1 mL × 2 | 10 mL |
Applications
- Hot-start PCR
- Taq-based qPCR
- TA cloning of PCR products
Storage & Handling
All components are stored at -20°C with a shelf life of two years.
Usage Notes
- Template purity is critical; impure templates should not exceed one tenth of the reaction volume.
- Primer working concentration is typically 0.2 µM.
- Set the annealing temperature about 5°C below the primer Tm.
- Use an extension time of about 1 kb/min.
- Excessive cycle numbers increase non-specific products.
- Thaw components and keep them on ice while assembling the reaction.
Safety & Regulatory
For research use only. Not intended for clinical, diagnostic or therapeutic use. Follow the manufacturer's manual and standard laboratory safety practice.
Customization & Add-ons: Need this molecular biology reagent tailored to your workflow? We can help with bulk or custom pack sizes, alternative formats (spin column, magnetic bead or reagent-only kits; premixed or stand-alone reference dye), matched extraction, cDNA synthesis and qPCR workflows, and sourcing of related enzymes, ladders and consumables not in our catalog. Click Talk to a Scientist to submit a request, email us at support@biohippo.com, or explore our Research Services — our team will follow up with feasibility details and next steps.
