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, MgCl(2) concentration, PCR cycling conditions, and ramp times were evaluated. RESULTS: Optimization of each parameter, including use of the OmniMix HS-lyophilized beads, 6 mM MgCl(2) 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.
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, MgCl(2) concentration, PCR cycling conditions, and ramp times were evaluated. RESULTS: Optimization of each parameter, including use of the OmniMix HS-lyophilized beads, 6 mM MgCl(2) 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.