A real-time fluorescence quantitative PCR instrument consists of a fluorescence quantification system and a CCD system. The CCD monitors fluorescence during the cycling process in real time. A computer connected to the real-time instrument collects the fluorescence data. The data is displayed graphically using developed real-time analysis software. Raw data is plotted as a graph of fluorescence intensity versus cycle number. Once the raw data is collected, analysis can begin. The real-time instrument software normalizes the collected data to compensate for differences in background fluorescence. After normalization, threshold levels can be set and analyzed for fluorescence data.
Real-time fluorescence quantitative PCR instruments are widely used in industries and fields such as basic medical research, pathogen detection, molecular cloning, genetic screening, gene expression, genotyping, transgenic testing, food safety testing, and public health epidemic monitoring.