| Field | Specification |
|---|---|
| Applications | |
| Source | Recombinant (E. coli) |
| Storage | |
| Shelf life | |
| Catalog no. (Mfr.) | |
| Main SKU |
Product Overview
Pfu DNA Polymerase is a thermostable protein isolated and purified from recombinant E.coli strains containing Pyrococcus furiosus DNA Polymerase gene, with a molecular weight of about 90 kDa. With 5'→3' polymerase activity and 3'→5' exonuclease activity, it can remove mismatched bases inserted during extension. Pfu DNA Polymerase is a high-fidelity thermostable polymerase. The PCR product is blunt-ended and can be directly linked to the blunt-ended cloning vector, or the end is treated with A and then connected to the TA vector.
The enzyme is supplied at 5 U/µL together with 10× Pfu Buffer and ddH₂O for 50 µL PCR reactions; dNTPs are not supplied. The enzyme can amplify single-copy genes from the human genome. A 50 µL reaction uses 0.5 µL of enzyme and 1 µL of dNTP mix and is cycled at 94°C for 3 min, then 30 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 5 min.
Specifications
| Concentration | 5 U/µL |
|---|---|
| Unit definition | 1 unit incorporates 10 nmol deoxynucleotides in 30 minutes at 74°C |
| Source | Recombinant (E. coli) |
| Activities | 5'→3' polymerase and 3'→5' exonuclease (proofreading) |
| Purity | Greater than 99% by SDS-PAGE |
| Molecular weight | About 90 kDa |
| Supplied buffer | 10× Pfu Buffer (dNTPs not supplied) |
| 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 | Blunt; can be A-tailed for TA vector cloning |
Kit Components
| Component | 50 µL | 100 µL |
|---|---|---|
| Pfu DNA Polymerase (5 U/µL) | 50 µL | 100 µL |
| 10× Pfu Buffer | 0.5 mL | 1 mL |
| ddH₂O | 1 mL | 1 mL × 2 |
Applications
- High-fidelity PCR amplification
- Amplification of single-copy genes from human genomic DNA
- Blunt-end cloning of PCR products
Storage & Handling
All components are stored at -20°C with a shelf life of two years.
Usage Notes
- dNTPs are not supplied and must be added separately; the 50 µL protocol adds 1 µL of dNTP mix, and the working concentration should be confirmed with the manufacturer before use.
- Add Pfu DNA Polymerase after the dNTPs to prevent exonuclease degradation of the dNTPs.
- Mix the buffer thoroughly before use.
- Template purity is critical; impurities should not exceed one tenth of the reaction volume.
- Dilute previously amplified products 1,000–10,000× before re-amplification.
- Assemble the reaction components on ice and centrifuge briefly before cycling.
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.
