Multiple Displacement Amplification - Materials - Phi 29 DNA Polymerase

Phi 29 DNA Polymerase

Bacteriophage Φ29 DNA polymerase is a high-processivity enzyme that can produce DNA amplicons of 7 to 10 kb length. Its high fidelity and 3’–5' proofreading activity reduces the amplification error rate to 1 in 106−107 bases compared to conventional Taq polymerase with a reported error rate of 1 in 9,000. The reaction can be carried out at a moderate isothermal condition of 30°C and therefore does not require a thermocycler. It has been actively used in cell-free cloning, which is the enzymatic method of amplifying DNA in vitro without cell culturing and DNA extraction. The large fragment of Bst DNA polymerase is also used in MDA, but Ф29 is generally preferred due to its sufficient product yield and proofreading activity.

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