title: Parameters involved in the conversion of real-time PCR assays from the ABI prism 7700 to the Cepheid SmartCycler II.
abstract: 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.
  
  
  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.
  
  
  CONCLUSIONS
  We conclude that careful consideration of several analytical parameters can result in a smooth transition of assays between real time PCR platforms.
authors.affiliation: Department of Pathology, Dartmouth Medical School, Dartmouth-Hitchcock Medical Center, One Medical Center Drive, Lebanon, NH 03756, USA.
authors.forename: Heather A
authors.initials: HA
authors.lastname: Bentley
authors.forename: Dorothy R
authors.initials: DR
authors.lastname: Belloni
authors.forename: Gregory J
authors.initials: GJ
authors.lastname: Tsongalis
mesh_terms: D007202:Indicators and Reagents; D015636:Magnesium Chloride; D016133:Polymerase Chain Reaction / Q000295:instrumentation* / Q000379:methods / Q000592:standards*; D013696:Temperature; D013995:Time
chemical_list: D007202:Indicators and Reagents; D015636:Magnesium Chloride