Parameters involved in the conversion of real-time PCR assays from the ABI prism 7700 to the Cepheid SmartCycler® II

Objective: We examined several critical parameters that must be optimized when converting between the ABI Prism 7700 real-time PCR platform and the Cepheid SmartCycler® II while using the same primer and probe sequences. Design and methods: A lyophilized master mix, MgCl2 concentration, PCR cycling conditions, and ramp times were evaluated. Results: Optimization of each parameter, including use of the OmniMix™ HS-lyophilized beads, 6 mM MgCl2 concentration, changes in PCR cycling parameters, and increased ramp time were necessary to convert this real time PCR assay to a new platform. Conclusion: We conclude that careful consideration of several analytical parameters can result in a smooth transition of assays between real time PCR platforms.