How does Real-Time PCR work?

Real-time PCR or qPCR is a powerful technique that combines DNA amplification and detection into a single step. It allows for the real-time monitoring of the amplification process by measuring the accumulation of amplification products in real time throughout the PCR process as it occurs, with product quantification after each cycle.

The real-time monitoring is achieved by adding a fluorescent reporter molecule in each reaction well, which produces an increasing amount of fluorescence as the DNA amplification progresses. Specialized thermal cyclers equipped with fluorescence detection modules enable detection of the fluorescence signals.

The amount of fluorescence emitted is directly proportional to the total amount of amplicon produced. The change in fluorescence over time is used to calculate the amount of amplicon generated in each cycle. 

Real-time monitoring allows for more accurate and sensitive quantification of DNA compared to traditional PCR methods.