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The maximum cycle threshold (Ct) value was set at 40.
Analysis of the training set allowed the identification of the threshold value for data analysis (0.2); the maximum cycle threshold (Ct = 37); and the cut-off ΔCt value (7) for the EGFR TMDA.
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The maximum CT (cycle threshold) value was set to 40.0 for all analyses.
Maximum cycle number (MCN) or ideal fitness threshold (IFT) could be set for terminating algorithm.
Using the Roche Lightcycler software, SYBR Green fluorescence measurements were plotted relative to cycle number and the 2nd derivative maximum method was used to determine the cycle threshold values (Ct) for each sample.
*MLE, maximum-likelihood estimation; CI, confidence interval; Ct, cycle threshold; qRT-PCR, quantitative reverse transcription PCR; –, not applicable.
For quantification, the cycle threshold number (Ct) exhibiting the maximum curve growth rate was determined.
*Ct, cycle threshold; LACV, La Crosse virus; MLE, maximum-likelihood estimate of the proportion of infected mosquitoes; RT-PCR, reverse transcription PCR.
Quantification was achieved using Cycle Threshold (CT) measured with the second derivative maximum method (LightCycler 480 Software version 1.5.0.39; Roche Applied Science).
The second derivative maximum algorithm included within the instrument software was used to determine cycle threshold (Ct) values for each region.
The cycle threshold Cp was automatically calculated in second derivative maximum method, and the expression of each gene relative to three housekeeping genes was calculated as ΔCp.
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