In 1993, Higuchi et al(1) demonstrated real-time or kinetic monitoring of DNA amplification. They used a conventional PCR system rigged with an UltraViolet lamp to irradiate PCR samples containing ethidium bromide and a camera to record the emission. Their technique revealed that scientists could monitor the accumulation of double-stranded DNA during PCR by measuring the increase in fluorescent emission as ethidium bromide bound to PCR products. The critical revelation was that a linear relationship was established between the amount of target DNA and the amount of PCR product generated by amplification after a specific number of amplification cycles. This technique was developed to become real-time or quantitative PCR (qPCR). QPCR and RT-qPCR are now the techniques of choice for identification and quantification of genomic and transcriptomic sequences. They are used to measure gemomic sequences, viral load or bacterial pathogens in clinical samples, RNA (via cDNA) copy numbers to determine the level of gene product in a tissue sample, to verify microarray or RNAi data. It is also widely used in a number of biotechnology applications, including the quantification of foreign genes in genetically modified foods and for forensic sample identification such as to identify victims of the 9/11 tragedies in New York. Since Higuchi’s initial experiments, qPCR methodologies have improved with specific detection chemistries and integrated instrumentation. 1Higuchi, R. et al. (1993) Kinetic PCR analysis: real-time monitoring of DNA amplification reactions. Biotechnology 11, 1026