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welcome to the applied biosystems tech talk video series where we break down real-time pcr to make it easier for you are you using applied biosystems cyber green chemistry for your real-time experiment and getting multiple peaks in your melt curve and are you asking yourself is this a bad thing letamp;#39;s get technical and start with a little background the dye in cyber green master mix is free floating hereamp;#39;s how it works when cyber green is just swimming around in the tube it doesnamp;#39;t give off much fluorescence even when we zap it with a light source on a real-time instrument but cyber green dye really likes to bind to double-stranded dna when it does and you hit it with light the dye gets excited and fluoresces at least in theory the basic idea is this as pcr creates more and more product the signal from cyber green dye goes up proportionally in practice though this doesnamp;#39;t always happen thatamp;#39;s because cyber green dye binds to any do