TempliPhi™, based on Phi29 DNA polymerase, is used to exponentially amplify single- or double-stranded circular DNA by rolling circle amplification (RCA).
Prepare templates for cycle sequencing, cloning, and transformation from circular DNA starting material efficiently, in 4 to 6 hours
Generate microgram quantities of template DNA from picogram amounts of circular DNA
Use amplified DNA directly for cycle sequencing without purification
Amplify DNA from bacterial or M13 liquid cultures, colonies, plaques, glycerol stocks, or purified circular (plasmid or M13) DNA
Reduces time, labor, and quantity of consumables needed for sequencing template preparation
Allows easy automation of workflow for efficient low- to medium-throughput small DNA vector preparation
TempliPhi™ kits use a unique kind of circular DNA amplification to efficiently prepare micrograms of DNA from picogram circular DNA starting material. TempliPhi™ kits use isothermal rolling circle amplification (RCA) for the exponential amplification of circular DNA using bacteriophage Phi29 DNA polymerase. The amplification used is often called multiple strand displacement amplification, or MDA. Phi29 DNA polymerase is active at 30°C, and isothermal amplification is performed at this temperature without the need for thermal cycling. The TempliPhi™ protocol allows amplification of 96 samples from bacterial colonies with less than 20 min of hands-on time.
Two different types of TempliPhi™ DNA Amplification Kits are available to meet different throughput requirements. TempliPhi™ DNA Amplification Kit for 2000 reactions, designed for automation, contains pre-mixed components and requires overnight incubation. TempliPhi™ 100 and TempliPhi™ 500 reaction kits contain separate, unmixed components for enhanced stability and preparations are ready in 4 to 6 hours.
