phi29 Highlights

phi29 DNA Polymerase exhibits strong strand displacement activity and high processivity to enable efficient isothermal DNA amplification from low DNA template amounts. Its 3′ → 5′ exonuclease activity delivers high fidelity amplification, making it an appropriate solution for sequencing DNA template preparation.

  • Strong strand displacement activity facilitates isothermal amplification
  • Strong processivity delivers products up to 70 kb in length
  • High-fidelity amplification supports DNA template preparation for sequencing
  • Custom formats available, including high concentration
  • Highly stringent enzyme production ensures quality performance across lots


  • Multiple displacement amplification (MDA)
  • Rolling circle amplification (RCA)
  • Whole genome amplification (WGA)
  • Cell-free cloning
  • Preparation of DNA template for sequencing

Rolling circle amplification diagram

Figure 1.
Schematic of rolling circle amplification using phi29 DNA polymerase.

QC Specifications
Description Specification
Protein Purity Assay ≥ 99%
Exonuclease Assay Functional
Specific Activity 100,000 – 190,000 U/mg
DNA contamination Assay (E. coli, mammalian, library)* < 10 copies
Phosphatase Contamination Assay* < 1% released
Endonuclease Contamination Assay* Not detectable

*As assessed using 1,000 U of enzyme input per assay.


Unit definition: One unit is defined as the amount of enzyme required to convert 50 pmol of dNTPs into a polynucleotide fraction in 10 minutes at 30°C

Reaction conditions:
1X phi29 Pol Reaction Buffer
Incubate at 30°C

Storage Buffer: 10 mM Tris-HCl, pH 7.4, 0.1 mM EDTA, 1 mM DTT, 100 mM KCl, 50% Glycerol, 0.5% Tween
1X phi29 Pol Reaction Buffer: 33 mM Tris-acetate (pH 8.2 at 25°C), 10 mM Mg-acetate, 66 mM K-acetate, 0.1% (v/v) Tween 20, 1 mM DTT

Heat inactivation: 65°C for 10 min

Molecular weight: 66.3 kDa
5’ – 3’ Exonuclease activity: No
3’ – 5’ Exonuclease activity: Yes
Strand Displacement activity: High
Fidelity: High
Processivity: High