phi29 DNA Polymerase is the replicative polymerase from the Bacillus subtilis phage phi29 (Φ29). This polymerase is a highly processive polymerase (up to more than 70 kb) featuring strong strand displacement activity, which allows for highly efficient isothermal DNA amplification. phi29 DNA Polymerase also possesses an inherent 3´→ 5' proofreading exonuclease activity, thus the fidelity of the synthesized DNA fragments is very high. 3'-modified primers are highly recommended to reduce the cleavage effect of the exonuclease activity on the primer.
We also offer phi29 DNA Polymerase in lyophilized form, please contact us for more details.
Strong strand displacement capability
Continuous synthesis capability
High fidelity of amplification
DNA isothermal rolling loop amplification
DNA synthesis requiring strong strand displacement
Rapid replication of plasmids
Rapid replication requiring high fidelity
Stored at -20°C for 3 years.
1 x phi29 Buffer: 50 mM Tris-HCl pH 7.5, 10 mM MgCl2, 10 mM (NH4) 2SO4, 4 mM DTT. If necessary, the final concentration of 1 mM DTT and 0.2 mg/ml BSA can be added separately to improve the reaction efficiency.
The optimum reaction temperature of the enzyme is 30℃.
The enzyme can be inactivated at 65℃ in 10 min.
Adjust the concentration of dNTP from 100 to 500 μM according to the type of experiments.
The addition of Yeast Pyrophosphatase to the reaction mixture with phi29 DNA Polymerase may enhance DNA synthesis.
Thio-modification at the 3' end of the primers can avoid primer degradation.
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