| Description |
φ29 DNA Polymerase is a highly processive polymerase from Bacillus Subtilis phage φ29. It has exceptional strand displacement and processive synthesis properties, which allows for highly efficient isothermal DNA amplification. The enzyme possesses a 3'→5' exonuclease (proofreading) activity acting preferentially on single-stranded DNA, allowing high-fidelity DNA synthesis. |
| Applications |
- Rolling circle amplification (RCA): generation of periodic DNA nanotemplates
- Multiple displacement amplification (MDA)
- Unbiased amplification of whole genome (WGA):
- amplification of DNA for SNP and STR detection
- cell-free amplification of DNA from single cells
- pathogenic organisms or metagenomes
- amplification of DNA from filter paper blood spot samples
- DNA template preparation for sequencing
- Protein-primed DNA amplification
- In situ genotyping with padlock probes
- Recombination based-cloning
- Cell-free cloning of lethal DNA
- RNA-primed DNA amplification
|
| Source |
A recombinant E. coli strain carrying the phi29 DNA Polymerase gene from bacteriophage phi29 |
| Unit Definition |
One unit is defined as the amount of enzyme that will incorporate 0.5 pmol of dNTP into acid insoluble material in 10 minutes at 30°C. |
| Components |
- phi29 DNA Polymerase: 100,000 U/ml in 10 mM Tris-HCl, 100 mM KCl, 0.1 mM EDTA, 1 mM DTT, 0.5% Tween-20, 0.5% NP-40, 50% glycerol, pH 7.4 @ 25°C
- 10X Reaction Buffer: 500 mM Tris-HCl, 100 mM (NH4)2SO4, 40 mM DTT, 100 mM MgCl2, pH 7.5 @ 25°C
|
| Quality Control |
The absence of endodeoxyribonucleases confirmed by appropriate quality tests. |
| Storage Condition |
-20 °C |