Phi29 DNA Polymerase

Description

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. 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. The enzyme can be heat-inactivated, tolerates dUTP and produces blunt-ended DNA.

Benefits:

• Isothermal amplification
• Extreme processivity
• High fidelity

Area of applications:

• Isothermal DNA amplification for sequencing or cloning
• Rolling circle amplification (RCA)
• Multiple displacement amplification (MDA)
• Amplification of DNA for SNP and STR detection
• Protein primed or RNA primed DNA amplification
• Whole Genome Amplification (WGA)

Concentration:

• 10 U/μl (5-100 U/μl upon request)

Storage and dilution buffer:

• Glycerol buffer: 20 mM Tris-HCl (pH 7.5), 0.1 mM EDTA, 100 mM KCl, 0.5% Tween-20, 2 mM TCEP, 50% glycerol
• Glycerol-free buffer: 20 mM Tris HCl (pH 7.5), 1 mM EDTA, 100 mM KCl, 750 mM trehalose, 5 mM TCEP

Storage and transportation conditions:

• From -24°С to -16°С – 1 year
• Avoid of freezing/thawing cycles