Figure 2
From: Model based analysis of real-time PCR data from DNA binding dye protocols
Effect of CT on the initial template amount estimation. (A) Product yield vs. cycle number for the amplification of three serial dilutions (0.1, 1 and 10) of cDNA from mouse midbrain with β-actin specific primers performed in triplicate. Horizontal lines show the values for the product yield at which CT was calculated. (B) Ratios between each T0 determination and T0 estimated for dilution 1 at the smallest CT value. T0 was calculated assuming constant amplification efficiency and using different amounts of PCR product for the estimation of CT values. Data points are the mean for triplicates. (C) Relative error for the quantification of data presented in A. Bars are the relative error of quantification as percentage (mean ± SEM for triplicates). (D) Real-time PCR amplification of the same mouse midbrain cDNA sample with β2 microglobulin specific primers using 0.1 and 0.25 units of Taq DNA polymerase. Horizontal lines show the values for the product yield at which CT was calculated. (E) Relative error for the quantification of data presented in D. Bars are the relative error of quantification as percentage (mean ± SEM for triplicates).