phi29 DNA Polymerase
Rapidly obtain high amplification yields from trace amounts of DNA without the use of a thermal cycler. With extremely high processivity, and high fidelity due to the presence of a 3’-5’ exonuclease, phi29 DNA Polymerase enables efficient synthesis of long DNA fragments with a minimal number of errors or amplification bias, making it ideal for applications such as rolling circle amplification (RCA), whole-genome amplification (WGA), single-cell sequencing, viral genome identification, and cell-free cloning.
Rapidly obtain high amplification yields from trace amounts of DNA without the use of a thermal cycler. With extremely high processivity, and high fidelity due to the presence of a 3′-5′ exonuclease, phi29 DNA Polymerase enables efficient synthesis of long DNA fragments with a minimal number of errors or amplification bias, making it ideal for applications such as rolling circle amplification (RCA), whole-genome amplification (WGA), single-cell sequencing, viral genome identification, and cell-free cloning.
For many applications, using traditional PCR-based reactions requires potentially difficult optimization. However, phi29 DNA Polymerase allows you to skip this optimization with a simple reaction setup and a constant reaction temperature of 30°C for DNA amplification.
Amplification reactions with phi29 DNA Polymerase only require random hexamers for polymerase binding. However, due to the strong 3′-5′ exonuclease activity of phi29 DNA Polymerase, unmodified hexamers are prone to degradation by phi29 DNA Polymerase, and the use of unmodified hexamers in the reaction may lead to inefficient DNA amplification. Therefore, we recommend random hexamers modified with 3′ phosphorothioate, such as 3′ Phosphorothioate-modified Random Hexamer (Cat. # 3807), which are resistant to 3′-5′ exonuclease activity.
Overview
- Obtain high-yield, isothermal amplification from as little as 5 pg of DNA
- Get accurate amplification from long DNA templates
- Streamline protocols by skipping the need for optimization of PCR primer and cycling conditions
Achieve higher DNA yields with phi29 DNA Polymerase from Takara Bio
Amplification reactions were performed using phi29 DNA Polymerase from Takara Bio or phi29 DNA Polymerase from competitor companies, with 0.5 ng pUC19 plasmid DNA. Each reaction was incubated at 30°C. An additional reaction using Competitor C’s mutant phi29 polymerase was incubated at 42°C. The DNA yield was measured at 2 and 4 hr. The phi29 DNA Polymerase from Takara Bio yielded the highest amount of DNA compared to other phi29 DNA polymerases from competitors. The yield was also comparable to that of the mutant phi29 DNA polymerase from Competitor C.
More Information
Source: E. coli expressing Bacillus subtilis phage phi29
Applications
- Rolling circle amplification
- Multiple displacement amplification
- Whole-genome amplification
- Cell-free cloning
- Viral genome amplification
- Preparation of DNA from single cells for sequencing
Additional production information
Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Product Components List to determine kit components. Certificates of Analysis and Product Components Lists are located under the Documents tab.
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