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Figure 1 | BMC Molecular Biology

Figure 1

From: Quantitative polymerase chain reaction analysis by deconvolution of internal standard

Figure 1

Determination of initial copy number of RCAN1.1 control DNA by efficiency-independent qPCR. (a) Linear correlations of c q vs. log10(f i U j + N i ) are shown as a function of the assumed template copy number (U j , on a logarithmic axis), and fluorescence threshold (T k ). Colors are keyed to indicate 0.01 unit steps of R2, and the optimal correlation (R2 = 0.995, U = 9.91 × 105 copies, T = 1.14 × 104 fluorescence units) was identified by iteration over 100 estimates (5 × 105U j ≤ 1 × 107), and 50 thresholds (104T k ≤ 106) where R max = 2.47 × 106 fluorescence units. (b) Graph of c q vs. log10(f i U + N i ) representing the optimal estimate U from the same experiment, which was conducted using 10 samples (replicate dilutions of f i = 1.0, 0.5, 0.25, 0.125, and 0.0625, with N i = 2.5 × 105 amplicon copies added to four samples, as indicated by the dashed lines. Graph line: c q = -3.53 log10(f i U + N i ) + 37.6 (R2 = 0.995).

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