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. 2021 May 29;11(6):496. doi: 10.3390/life11060496

Figure 6.

Figure 6

Effect of pipetting error (solid lines, Equation (15)) and Poisson sampling variation (dotted lines, Equation (16)) on the number of copies in the reaction and the observed Cq values. (A) Actual number of target copies in the reaction. Below an average of four copies (vertical dotted red line), an increasing fraction of the reactions be false negative reactions (pink area). (B) Range of observed Cq values calculated for an efficiency value of 1.9 and a Cq of 35 for an average input of 10 copies. Note that the lower Cq values come from reactions with the high number of target copies (panel A). (C) Range of Cq values for different average number of target copies in the reaction (Equation (17)). The graphs are plotted for a PCR efficiency of 1.9; the effect of the PCR efficiency on the observed Cq values is illustrated in Figure 5 and Figure 8.