Figure 2From: Quantitative polymerase chain reaction analysis by deconvolution of internal standardEstimated vs. actual template copy numbers, applying the same data to four methods of analysis. Symbols are mean values for each amplicon (n = 3, 10 samples each as in Figure 1; y-axis error bars, minimum and maximum copy number estimates; x-axis error bars, 10% s.d. from a priori error in pipetting and dilution). (a) Efficiency-independent method described in this paper (absolute quantification). RCAN1.1 dashed line (y = 1.05x - 5180, R2 = 0.9996), β-actin dotted line (y = 1.00x - 83, R2 = 0.9999). (b) Linear Regression of Efficiency (LRE) method (absolute quantification). Average correlations of fluorescence data to sigmoidal curves: R2 = 0.9987 (n = 30, RCAN1.1, Emax = 0.912 ± 0.007 s.d.) and R2 = 0.9999 (n = 30, β-actin, Emax = 0.941 ± 0.007 s.d.). RCAN1.1 dashed line (y = 0.618x - 4790, R2 = 0.9903), β-actin dotted line (y = 0.240x - 5650, R2 = 0.9751). (c) LinReg method (relative quantification). Y-axis is scaled to the geometric mean of data (open circle). Average correlations (log10R n vs. n, over 4 cycles) for determination of Φ: R2 = 1.000 (n = 30, RCAN1.1) and R2 = 1.000 (n = 30, β-actin). RCAN1.1 dashed line (y = 1.28x - 0.0951, R2 = 0.9601), β-actin dotted line (y = 0.825x - 0.0761, R2 = 0.9880). (d) Serial dilution method (relative quantification). Y-axis is scaled to the geometric mean of data (open circle). Average graph correlations (c q vs. log10A 0 ) for determination of Ψ: R2 = 0.9959 (RCAN1.1) and R2 = 0.9947 (β-actin). RCAN1.1 dashed line (y = 1.19x - 0.261, R2 = 0.9888), β-actin dotted line (y = 0.965x - 0.048, R2 = 0.9820).Back to article page