There are various methods of determining amplification efficiency as summarized under number 1 and 2 and in the bullets below:
1. From calibration curve slope as determined by:
· Fit-point method;
· Second derivative maximum for the 4 parametric logistical model;
2. From single amplification plots using algorithms like:
· Mid-value point regression — also known as data analysis for real-time PCR or DART-PCR;
· Window-of linearity algorithm or LinREG PCR; and
· Noise-resistant iterative nonlinear regression or RealTime PCR Miner.
So, I would like to know the most rigorous and classical method examines the slope of a calibration curve?
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