Twenty years of qPCR: a mature technology?

Quantitative PCR is the "gold standard" technology to quantify nucleic acids and, since the first report describing real-time PCR detection in 1993, its use has been grown exponentially. More recent technological advancements have extended the field of applications ranging from high-resolution melting detection to digital PCR. Nowadays, it is a very accessible technique, but some pitfalls should be overcome in order to achieve robust and reliable analysis.