Figure 1
Amplification efficiency determination via standard curve analysis. (A) Screenshot of the amplification profiles generated by five quantities of lambda gDNA, ranging from 188,000 to 19 genomes in 10-fold increments. The fluorescence threshold is represented by the red line spanning the lower region of the profiles. Note that each amplification profile was produced by averaging the fluorescence readings of three replicate amplification reactions. (B) Plotting Log target quantity vs. Ct generates a line in which the amplification efficiency is derived from the slope (Eslope) and the number of amplicon molecules at threshold (Nt) is derived from the intercept via linear regression analysis [2]. fg Lambda: femtograms of lambda gDNA, N0: the number of lambda genomes, Ct: the threshold cycle, r2: linear regression correlation coefficient, Ft: the fluorescence threshold, FU: fluorescence units