What is real-time PCR?
Real-time PCR is a variation of endpoint PCR that allows amplification to be monitored in real time using specially designed instruments that perform both the thermal cycling steps to amplify the target and fluorescence detection to monitor changes in fluorescence in real time. Fluorescently labeled oligonucleotide probes or primers or DNA-binding fluorescent dyes such as SYBR Green are used to detect and quantitate the PCR product throughout each PCR cycle. As the PCR product accumulates, fluorescence of the probe, primer or DNA-binding dye increases or, with some technologies, decreases. During the exponential phase of PCR, the change in fluorescence is proportional to the accumulation of PCR product. Real-time PCR is commonly used to quantitate a specific DNA target or, in the case of real-time RT-PCR, an RNA target. For more information, see the enotes article Your Key to Real-Time Quantitative PCR and the Nucleic Acid Amplification chapter of the Promega Protocols and Applications
