phi29 DNA Polymerase

Opis towaru

Recombinant phi29 DNA Polymerase is a classical enzyme dedicated for use in common isothermal DNA amplification applications

that are carried out at moderate temperature based on a strand displacement activity.

The enzyme is supplied with an optimized high-performance buffer. The user has to add dNTPs, template and primers.

phi29 DNA Polymerase was discovered and characterized by M. Salas.

The Polymerase has strong strand displacement activity and efficient 5’-3’ polymerase activity working at about 4 - 35°C

and synthesizing DNA from minor amounts to enormous yield up to visibly increased the viscosity of the reaction mixture.

The enzyme has no 5’-3’ exonuclease activity, but has strong 3’ - 5’ exonuclease (proofreading) activity, and may degrade primers,

therefore the use of 3’ protected exo-resistant primers is recommended in the literature. The enzyme can be heat-inactivated,

tolerates dUTP and produces blunt-ended DNA.

Most important technical characteristics of phi29 DNA Polymerase (based on abundantly available scientific literature):

Strong strand displacement activity.

5’ - 3’ polymerase activity on DNA templates (and on RNA templates).

Strong 3’ - 5’ (proofreading) exonuclease activity, may degrade unprotected primers.

No 5’ - 3’ exonuclease activity.

Optimal reaction temperature is 30 °C.

Working temperature range is 4 - 35 °C, depends on application.

Optimal reaction time depends on application.

The enzyme is inactivated in 10 minutes at 65 °C.

Can extend both DNA and RNA primers.

Addition of pyrophosphatase may accelerate the reaction.

The use of this product in certain applications, some additives or protocols may be covered by patents.

The user has to analyse all applicable Limited Use Label Licenses and may need licensing for certain cases.

Applications

Isothermal DNA amplification for sequencing, cloning

RCA – rolling cycle amplification

MDA – multiple displacement amplification

WGA - whole genome amplification

Protein - primed or RNA-primed DNA amplification

Benefits

High yield DNA amplification at constant 30°C temperature

Robust polymerase with strong strand displacement activity

High processivity, synthesis of over 70 kb long DNA strains

High fidelity, low error rate for sequencing and cloning

Classical enzyme/buffer formulation for known phi29 applications