The NxGen Phi29 DNA Polymerase sources its enzyme from a recombinant E. coli strain carrying the phi29 DNA polymerase gene from bacteriophage phi29.
1 unit is defined as the amount of polymerase required to convert 0.5 pmol of dNTP's into acid insoluble material in 10 minutes at 30 °C.
The enzyme storage buffer consists of 10 mM Tris-HCl, 100 mM KCl, 0.1 mM EDTA, 1 mM dithiothreitol, 0.5% Tween-20, 0.5% NP-40, 50% glycerol, pH 7.4 @ 25 °C and the 10X phi29 DNA Polymerase Buffer is composed of 500 mM Tris-HCl, 100 mM (NH4)2SO4, 40 mM Dithiothreitol, 100 mM MgCl2, pH 7.5 @ 25 °C.
NxGen Phi29 DNA Polymerase is a highly processive and high-fidelity DNA Polymerase that enables accurate amplification of DNA from limited or degraded samples.
Powerful strand displacement activity, making it ideal for challenging samples or damaged DNA
Highly processive, allowing for amplification of up to 70,000 base insertions per binding event
High-fidelity 3'->5' proofreading exonuclease function for accurate DNA replication with low error rates
NxGen Phi29 DNA Polymerase is a highly processive, high-fidelity DNA polymerase that is derived from bacteriophage phi29.
It is an isothermal DNA polymerase with high strand displacement activity and a 3′→ 5′ proofreading exonuclease function that can amplify DNA with high specificity and accuracy, even from limited or degraded samples.
Ideal applications:
Rolling Circle Amplification (RCA)
Whole Genome Amplification (WGA)
Strand Displacement Activity
The high processivity of the enzyme allows for efficient amplification of long DNA fragments up to 70,000 base insertions per binding event, while the high-fidelity ensures that the amplified DNA is of high quality and accuracy.
Figure 1. Whole genome amplification as outlined in Protocol 2 of this manual. Five different lots of NxGen phi29 DNA Polymerase were used and duplicate NTC and (+) human genomic amplification reactions were set up. Amplification reactions were incubated for 16 hours at 30 °C and products were analysed in a 0.7% agarose gel and stained with ethidium bromide to visualise amplified DNA. The red arrows indicate NTC reactions with detectable background (input template independent) amplification.
Unit Definition: 1 unit is defined as the amount of polymerase required to convert 0.5 pmol of dNTP's into acid insoluble material in 10 minutes at 30°C.
Source: A recombinant E. coli strain carrying the phi29 DNA Polymerase gene from bacteriophage phi29.
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