Thermo Scientific phi29 DNA Polymerase is a highly processive polymerase (up to more than 70 kb) featuring strong strand displacement activity, which allows for highly efficient isothermal DNA amplification. phi29 DNA Polymerase also possesses a 3'→5' exonuclease (proofreading) activity acting preferentially on single-stranded DNA or RNA. Therefore 3'-modified primers are highly recommended.

Highlights

• Highest processivity and strand displacement activity among known DNA polymerases – more than 70 kb long DNA stretches can be synthesized
• Highly accurate DNA synthesis
• Extremely high yields of amplified DNA even from minute amounts of template
• Amplification products can be directly used in downstream applications (PCR, restriction digestion, SNP genotyping, etc.)

Applications

• Rolling circle amplification (RCA): generation of periodic DNA nanotemplates
• Multiple displacement amplification (MDA)
• Unbiased amplification of whole genome (WGA, see Figure 1 in Supporting Data):
• 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

Note

Addition of Pyrophosphatase to the reaction mixture with phi29 DNA Polymerase may enhance DNA synthesis.

Use of this enzyme in certain applications may be covered by patents and may require a license.