We demonstrated the principles of the chemistry by use of an essential target and selected mutations found within the CFTR gene recommended by leading medical organizations. Data from the Wisconsin State Laboratory of Hygiene confirmed that the chemistry is accurate. For example, there were no incorrect determinations, and the percentage of correct determinations was high. The data presented here were tabulated by use of a broad sample set that included every mutation possible within the panel targeted with the exception of the 3199del6. For 24 samples, the entire manual process takes ϳ3 h to obtain final results. An upgraded prototype version of the CFTR test where all mutations are analyzed in a single well takes ϳ2.5 h to complete analysis of 96 patient samples (validation data not yet available). To demonstrate the system in a fully automated mode, we elected to use the Tecan genesis liquid-handling system. The entire automated process also takes ϳ3 h from prepared sample to final read for an entire 96-well plate or 24 samples. We presented data from two possibilities (when automation started at the PCR step and when it started at the TSE step) because there may be a need to perform PCR offline.