| Field | Specification |
|---|---|
| Applications | |
| Source | Recombinant (E. coli) |
| Storage | |
| Shelf life | |
| Catalog no. (Mfr.) | |
| Main SKU |
Product Overview
Taq DNA Polymerase is a thermostable protein isolated and purified from a recombinant E.coli strain containing Thermus aquaticus DNA Polymerase gene, with a molecular weight of about 90 kDa. It has 5'→3' polymerase activity and double-strand specific 5'→3' exonuclease activity, but no 3'→5' exonuclease activity. The Taq DNA Polymerase PCR product is a 3' single A sticky end, which can be directly connected to the TA vector.
The enzyme is supplied at 5 U/µL together with 10× PCR Buffer (Mg²⁺ free), 25 mM MgCl₂ and ddH₂O for 50 µL PCR reactions. The final Mg²⁺ concentration in the reaction is 2 mM and can be adjusted between 2 and 5 mM. A 50 µL reaction uses 0.5 µL of enzyme and is cycled at 94°C for 5 min, then 35 cycles of 94°C for 30 sec, 50–60°C for 30 sec and 72°C for 30 sec/kb, with a final 72°C for 10 min.
Specifications
| Concentration | 5 U/µL |
|---|---|
| Unit definition | Incorporates 10 nmol dNTPs into acid-insoluble material in 30 min at 74°C |
| Source | Recombinant (E. coli) |
| Activities | 5'→3' polymerase and double-strand specific 5'→3' exonuclease; no 3'→5' exonuclease |
| Purity | Greater than 99% by SDS-PAGE |
| Molecular weight | About 90 kDa |
| Supplied buffers | 10× PCR Buffer (Mg²⁺ free) and 25 mM MgCl₂ |
| 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 | 3' single A overhang for direct TA vector cloning |
Kit Components
| Component | 500 U (100 µL) (bulk only) | 2500 U (500 µL) |
|---|---|---|
| Taq DNA Polymerase (5 U/µL) | 100 µL | 500 µL |
| 10× PCR Buffer (Mg²⁺ Free) | 1 mL | 5 mL |
| 25 mM MgCl₂ | 1 mL | 5 mL |
| ddH₂O | 1 mL × 2 | 10 mL |
Applications
- PCR amplification
- Amplification from genomic, λ and plasmid DNA templates
- 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; low-purity template should not exceed one tenth of the reaction volume (up to 5 µL per 50 µL).
- Dilute previously amplified products 1,000–10,000× before re-amplification.
- Primer working concentration is 0.2 µM, prepared from a 10 µM stock.
- Set the annealing temperature about 5°C below the primer Tm.
- The cycling program uses 30 sec/kb of extension, while the manual's notes quote 500 bp/min; confirm the extension time for long amplicons with the manufacturer.
- Thaw components on ice and mix by inverting before 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.
