Real-time PCR is a technique that combines amplification and detection in a single step by correlating the PCR product of each cycle with a fluorescence intensity signal. They consist of a thermal cycler coupled to an optical system, which monitors the signal of the fluorophores used to detect the amplified product. Because the fluorescence of the fluorophores increases as the product is amplified, the amplification and detection processes are combined in a single step.

This technique has important features such as high specificity, wide detection range and rapid visualisation of the product.

Real-time PCR systems include a thermal cycler and a unit capable of detecting fluorescent signals to monitor the progress of the amplification reaction, as well as software for data analysis.

The fluorophores used to follow DNA amplification during real-time PCR can be either DNA affinity (SYBR Green) or DNA fragment-specific probes (Taqman probes).

The QuantStudio™ 5 kit has filters for the following fluorophore spectra: blue (FAM™and SYBR™), green (VIC™), yellow (ABY™, NED™, and TAMRA™), orange (JUN™ and ROX™) and red (Cy®5 and MUSTANG PURPLE™).