Up to now only one software package can automatically determine the realtime PCR efficiency sample-by-sample. In the Rotor-Gene™ 3000 software package (Corbett Research), it is called the comparative quantification. Amplification rate is calculated on the basis of fluorescence increase in the PCR exponential phase. Further algorithms and methods are described in recent publications to estimate the real-time PCR efficiency. These can be grouped in direct and indirect methods. Direct methods are based on either a dilution method or a measurement of the relative fluorescence increase in the exponential phase. On the other hand, indirect methods are published, doing the efficiency calculation on basis of a fit to a mathematical model, like sigmoidal, logistic models or an exponential curve fitting (for details see http://efficiency.gene-quantification.info).
Was this article helpful?