DNA polymerases
Pfu proofreading Polymerase
Proofreading DNA polymerase from Pyrococcus furiosus for high-fidelity PCR and blunt-end cloning
- Cat. No.
- M3004
- Category
- DNA polymerases
- Formats
- 3 formats
- Manufacturer
A recombinant thermostable DNA polymerase from the hyperthermophilic archaeon Pyrococcus furiosus, expressed in E. coli, with 5'-3' polymerase and 3'-5' proofreading exonuclease activity. Compared with Taq it is about ten times more accurate and yields blunt-ended products that suit cloning and expression. It is supplied at 2.5 U/µL with a 10x reaction buffer containing MgSO4.
Key features
Proofreading
The 3'-5' exonuclease removes misincorporated nucleotides, for about ten times the accuracy of Taq.
Blunt-ended products
No 3' A addition and no 5'-3' exonuclease activity: suited to blunt-end cloning.
Thermostable and processive
Half-life at high temperature is far longer than that of Taq.
Works with modified nucleotides
Accepts dNTPs, ddNTPs and fluorescent dNTPs or ddNTPs.
Applications
- High-fidelity PCR
- Blunt-end cloning
- Amplification of sequences for expression
- Primer extension reactions
- Applications that need a low error rate
Technical details
| Concentration | 2.5 U/µL |
|---|---|
| Supplied with | 10x PCR buffer with MgSO4: 60 mM (NH4)2SO4, 200 mM Tris-HCl pH 8.8, 100 mM KCl, 20 mM MgSO4, 1% Triton X-100, 1% BSA |
| Activity | 5'-3' polymerase and 3'-5' exonuclease; no 5'-3' exonuclease activity |
| Fidelity | About 10 times higher than Taq |
| Product ends | Blunt; TA cloning requires an A-tailing step |
| Recommended extension | About 1 minute per 250 bp of target, roughly twice the Taq time |
| Amplicon length | Fragments up to about 4 kb; above 5 kb optimisation is usually needed |
| Unit definition | 1 U incorporates 10 nmol of dNTPs into acid-insoluble material in 30 minutes at 72 °C |
| Storage | -20 °C; up to 3 months at +2/+8 °C |
| Shipping | On wet ice; full activity for at least 2 weeks at +15/+30 °C |
| Stability | Until the expiry date on the label when stored at -20 °C |
| Quality control | Amplification efficiency and reproducibility; absence of exo- and endonuclease, RNase and protease activity; self-priming test on human genomic DNA |
What you receive
The product
Shipped under controlled temperature when the product requires it.
Lot certificate
Every lot comes with its certificate.
Tracked shipment
Follow your shipment live, from our warehouse to your lab.
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DNA polymerasesSuperSens Taq DNA Polymerase GreenCat. No. DB-1329Formats 3 formatsFrequently asked questions
Can I ask for formats other than those listed?
Yes: custom formats and OEM packs are available on request. Tell us the quantities you need.
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