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TempliPhi DNA amplification kits from GE Healthcare utilise bacteriophage φ29 DNA polymerase to exponentially amplify single- or double-stranded circular DNA templates by rolling circle amplification. Approximately 2 µg of DNA from can be amplified from picogram amounts of starting material eliminating the need for overnight cell culture and conventional plasmid or M13 DNA purification.

Bacterial colonies can be picked from agar plates and added directly to the TempliPhi reaction. Alternatively, microliter quantities of a saturated bacterial culture or a glycerol stock can serve as starting material. Depending on the source of starting material, amplification is completed in 4–18 hours at 30°C with no need for thermal cycling.

DNA amplified by the TempliPhi method can be used directly in cycle sequencing reactions without any purification. The reagent cost is approximately $1.65 per sample (based on 5ml reaction vol.).

TempliPHI™ company protocol

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