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NxGen phi29 DNA Polymerase

NxGen phi29 DNA Polymerase

Product Details

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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

NxGen Phi29 DNA Polymerase is a highly processive and high-fidelity DNA Polymerase that enables accurate amplification of DNA from limited or degraded samples. 

Key features

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  • 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

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Product information

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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