Problems with detection in the Plateau phase of PCR

The following three figures show the plateau affect on 96 replicates and a fivefold dilution series. As stated earlier, the plateau region is the end-point of the reaction and is representative of the amount of product that you would see on
Agarose Gels. The 96 replicates in the exponential phase are very tight in both the linear and logarithmic views.

In the logarithmic view, Figure 7, the plateau for each reaction seems to occur in the same place, but this is solely due to the log scaling of the plot. Figure 6 shows the same 96 replicates in linear view. The reactions show a clear
separation in the plateau phase; therefore, if the measurements were taken in the plateau phase, quantitation would be affected.



The 5-fold dilution series, seen in Figure 8, seems to plateau at the same place even though the exponential phase clearly shows a difference between the points along the dilution series. This reinforces the fact that if measurements
were taken at the plateau phase, the data would not truly represent the initial amounts of starting target material.
Real-Time chemistry provides fast, precise and accurate results. Real-Time PCR is designed to collect data as the reaction is proceeding, which is more accurate for DNA and RNA quantitation and does not require laborious post PCR
methods.

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