Quantification of DNA by Real-Time Polymerase Chain Reaction (PCR)

There are few differences between the experimental steps necessary for amplifying template DNA in a real-time thermocycler and a standard polymerase chain reaction (PCR). In real-time PCR, it is necessary, however, to optimize the concentration of primers and probe and to perform a standard curve. It is also important to consider the data analysis method that will be used.