Figure 1.

Representations of real-time PCR amplification curves. The three phases of the amplification reaction are shown either on a linear scale (panel A) or on a semi-log scale (panel B). Panel A represents a typical amplification curve, while panel B depicts amplification curves generated from serial dilutions of the same sample, either undiluted or diluted 10- or 1000-fold (indicated as 1, 0.1 or 0.001, respectively). During the lag phase (phase I), the fluorescence resulting from DNA amplification is undetectable above noise fluorescence in part A, while in part B, some data points take negative values and are not represented. This phase is used to evaluate the baseline "noise" of the PCR amplification. Exponential amplification of the DNA is detected in phase II (cycles 16 to 23, panel A). This phase of the amplification corresponds to the linear portion of the curve in panel B (closed circles). A threshold value is usually set by the user to cross the log linear portion of the curve, defining the threshold cycle value (Ct). Phase II is followed by a linear or plateau phase as reactants become exhausted (phase III). The inserted equations describe the dynamic of the amplification during phase II.