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The real-time fluorescence-based quantitative polymerase chain reaction (qPCR) has. become the benchmark technology for the detection of nucleic acids in every area of. microbiology, biomedical research, biotechnology and in forensic applications. Unlike. conventional (legacy) PCR, which is a qualitative end-point assay, qPCR allows accurate.
Introduction. Real-time PCR, also called quantitative PCR or qPCR, can provide a simple and elegant method for determining the amount of a target sequence or gene that is present in a sample. Its very simplicity can sometimes lead to problems by overlooking some of the critical factors that make it work.igure 1. Graphical representation of real-time PCR data. Rn is the fluorescence of the reporter dye divided by the fluorescence of a passive reference dye; i.e.,Rn is the reporter signal normalized to the fluorescence signal of ROXâ¢. (A) In this view, Rn is plotted against PCR cycle number.