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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 in a previous tech talk video we discussed assay efficiency and how to calculate it now letamp;#39;s talk about how high or low assay efficiency impacts real-time pcr data the aim of all real-time pcr assays is to be 100 efficient which means thereamp;#39;s an exact doubling of template in every cycle we previously found that this can be calculated using a dilution series and the following equation in order to use the delta delta ct method for relative quantification the efficiencies of the target and endogenous control assays must be approximately equal within 10 percent if their efficiencies differ by more than 10 percent you should be aware that unequal pcr efficiency will reduce the accuracy of your calculations when you evaluate the total changes the applied biosystems portfolio includes well over 20 million pre-designed assays that have been validated in