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Real-time detection of PCR products involves using fluorescent dyes or probes to detect specific double-stranded DNA sequences. Fluorescently-labeled probes measure specific DNA amounts, while dyes measure total DNA. In this tutorial, we focus on fluorescent dye-based detection, where a dye emits fluorescence when bound to double-stranded DNA. The signal intensifies with each PCR cycle as more DNA is generated, with SyBr Green being a commonly used dye. SyBr Green binds to double-stranded DNA, emitting fluorescence as DNA quantity increases during PCR.
