Constructing a Standard Curve for Real-Time Polymerase Chain Reaction (PCR) Experiments.

It is essential to prepare a standard curve for every real-time polymerase chain reaction (PCR) experiment. This protocol is used to construct a standard curve in which the template concentration is unknown. Such a standard curve is suitable for optimization experiments and for performing relative quantification by the standard curve method. To construct a standard curve for absolute quantification, the same principles apply as those presented here, but the concentration of the standards must be determined by an independent method.