Method Overview - PCR

By PCR (polymerase chain reaction) a DNA or cDNA template is amplified more than thousandfold. This is achieved by an initial denaturation step at 94 °C or higher to separate the intertwined DNA strands, followed by primer annealing at 40 °C – 60 °C. In the final extension step, the temperature is raised again, and the DNA polymerase extends the primers, synthesizing the new strand. Denaturation, annealing, and elongation constitute a single PCR cycle where the DNA is doubled.

Thermocyclers (e.g. QuantStudio 5 Real Time PCR system (Thermo Fisher Scientific) or CFX96 Touch Real-Time PCR Detection System (Bio-Rad) at VelaLabs) provide the temperature protocol for the reaction by heating and cooling the sample.