phi29 DNA Polymerase

phi29 DNA Polymerase

phi29 DNA Polymerase is a highly processive recombinant polymerase with exceptional strand displacement activity, which allows for highly efficient isothermal DNA amplification. The enzyme is capable of up to 70 thousands base insertions per binding event. The polymerase also possesses a 3’to 5’ proofreading exonuclease activity acting preferentially on ssDNA or RNA. Therefore 3′-modified primers are recommended.

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Description

phi29 DNA Polymerase is a highly processive recombinant polymerase with exceptional strand displacement activity, which allows for highly efficient isothermal DNA amplification. The enzyme is capable of up to 70 thousands base insertions per binding event. The polymerase also possesses a 3’to 5’ proofreading exonuclease activity acting preferentially on ssDNA or RNA. Therefore 3′-modified primers are recommended.

Features

  • Recombinant polymerase derived from the Bacillus subtilis phage phi29 and over-expressed in E. coli.
  • Extremely processive polymerase, enabling amplification up to 70 kbp.
  • Extremely high yields of amplified DNA can be obtained even from minute amounts of template.
  • High-fidelity polymerase – possesses a 3’-5’ exonuclease (proofreading) activity acting preferentially on ssDNA or RNA.
  • Isothermal polymerase – no need for thermal cycling.
  • Due to heat lability of the enzyme, it is possible to use the amplification products directly in the downstream applications.

Applications

  • Rolling Circle Amplification (RCA).
  • In situ genotyping with padlock probes.
  • Amplification of DNA for SNP and STR detection.
  • Unbiased amplification of whole genome.
  • DNA template preparation for sequencing.
  • RNA-primed DNA amplification.
  • Multiple displacement amplification (MDA).
  • Cell-free amplification of DNA from single cells.
  • Amplification of DNA from environmental samples.

Unit definition

One unit of phi29 DNA Polymerase is defined as the amount of enzyme that will incorporate 0.5 pmol of dCMP into a polynucleotide fraction in 10 minutes at 30°C under standard assay conditions.

Literature

phi29 DNA polymerase user manual