While kinetic RT-PCR has a tremendous potential for analytical and quantitative applications, a comprehensive understanding of its underlying principles is important. Fidelity of real-time RT-PCR is associated with its "true" specificity, sensitivity, reproducibility, and robustness and, as a fully reliable quantitative method, it suffers from the problems inherent in RT and PCR, e.g., amplification of unspecific products, primer-dimers, amplification efficiencies, hetero-duplex formation, etc.13