Thermo Scientific phi29 DNA Polymerase is a highly processive polymerase featuring strong strand displacement activity, which allows for highly efficient isothermal DNA amplification. Produced from E. coli cells with a cloned gene 2 of Bacillus subtilis phage phi29, phi29 DNA Polymerase possesses 3'→5' proofreading exonuclease activity that acts preferentially on single-stranded DNA or RNA.
Features of phi29 DNA Polymerase
• Excellent processivity and strand displacement activity—more than 70 kb long DNA stretches can be synthesized
• Highly accurate DNA synthesis
• High yields of amplified DNA even from small amounts of template
Phi29 DNA Polymerase may be used in a wide variety of applications including:
• Rolling circle amplification (RCA)—generation of periodic DNA nanotemplates
• Multiple displacement amplification (MDA)
• Unbiased amplification of whole genome (WGA) (see figure below)
• Amplification of DNA for SNP and STR detection
• Cell-free amplification of DNA from single cells, pathogenic organisms, or metagenomes
• DNA template preparation for sequencing
• Protein-primed DNA amplification
• Recombination based-cloning
• Cell-free cloning of lethal DNA
• RNA-primed DNA amplification
1. The addition of pyrophosphatase to the reaction mixture with phi29 DNA Polymerase may enhance DNA synthesis.
2. Because of the enzymes’s 3’→5’ proofreading exonuclease, 3'-modified primers are highly recommended.