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Validation of the 2−ΔΔCt method. ΔCt calculated as Ct ,target gene − Ct ,18 s for each cDNA dilution. Data were fit using a least-squares regression analysis (with each reaction run in triplicate) to calculate the slope of the best fit line. A slope < 0.1 indicates that amplification efficiencies are adequately similar to use the 2−ΔΔCt method [31].
