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Table 3 Comparison of the accuracy between Pairwise Efficiency and the standard calibration curve method based on a chosen standard

From: Pairwise efficiency: a new mathematical approach to qPCR data analysis increases the precision of the calibration curve assay

Wells Conc. Efficiency F0 Ratio (PE) Error (%) Ratio (Ct) Error (%)
A1-A6 100 ng 0.73130 0.00800 1 N/A 1 N/A
A7-A12 100 ng 0.76200 0.00780     
B1-B6 100 ng 0.77170 0.00660     
B7-B12 100 ng 0.77230 0.00710     
C1-C6 50 ng 0.83530 0.00280 2.513 20% 2.47 19%
C7-C12 50 ng 0.79550 0.00290     
D1-D6 50 ng 0.81870 0.00290     
D7-D12 50 ng 0.82390 0.00300     
E1-E6 12 ng 0.75780 0.00060 8.519 6% 12.73 37%
E7-E12 12 ng 0.68420 0.00110     
F1-F6 12 ng 0.72470 0.00090     
F7-F12 12 ng 0.70420 0.00100     
G1-G6 3 ng 0.76180 0.00020 35.455 10% 57.41 44%
G7-G12 3 ng 0.66870 0.00020     
H1-H6 3 ng 0.72810 0.00020     
H7-H12 3 ng 0.66640 0.00020 Aver error: 12%   33%
  1. The efficiency of amplification of Actin beta was determined using Pairwise Efficiency or the standard calibration curve method (for standard method E values see Additional file 1: Table S3). The known dilution ratio (differences between DNA template concentrations) were used as a reference. 100 ng was taken as 1, and thus all diluted samples should have yielded the following values: 2 (for 50 ng), 8 (for 12 ng) and 32 (for 3 ng). The error values in determining the correct ratios were lower than those calculated by standard method. The average error for Pairwise Efficiency was equal to 12%, while the average error for standard method was equal to 33%