qPCR Analysis Software
Takes advantage of later cycles Formalized baseline adjustments Works even with high signal-to-noise ratioΓÇÖs
UCF researchers developed a simple mathematical model that accurately describes the entire PCR reaction profile using only two reaction variables that depict the maximum capacity of the reaction and feedback inhibition. This model allows quantification that is more accurate than existing methods and takes advantage of the brighter fluorescence signals from later cycles. Because the model describes the entire reaction, the influences of baseline adjustment errors, reaction efficiencies, template abundance, and signal loss per cycle could be formalized. We determined that the common cycle-threshold method of data analysis introduces unnecessary variance because of inappropriate baseline adjustments, dynamic reaction efficiency, and also a reliance on data with a low signal-to-noise ratio. An additional benefit of the global fitting method described here is that researchers can evaluate the quality of their raw reaction data against a idealized model to better address experimental artifacts.
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