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
One-tube, one-temperature format
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.
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