I know that the ideal amplification signal should be sigmoidal in shape, but how do I interpret non-ideal curves?
The package insert describes how to interpret the results. At a minimum, a curve representing PCR amplification must have a sharp rise in the positive direction. The point at which the sharpest rise occurs is called the inflection point. The inflection point corresponds to a second derivative peak that can be viewed by selecting the appropriate graph on the SmartCyler. Figure 1 below shows a low positive in two views, the primary curve and the second derivative plot. If you have technical questions or concerns regarding this test, contact us. Figure 1. Shown are the primary amplification curve and the second derivative view of the same curve.
