Amplification Plot

Amplification Plot. Model of a single amplification plot illustrating the nomenclature Ideally, this is within the range where the difference between cycles for different amplification plots remains constant Real-time PCR data are often represented in a graph called an amplification plot (Figure 1)

Amplification plot for Real TimePCR reaction for samples 2, 4, 6 and 8
Amplification plot for Real TimePCR reaction for samples 2, 4, 6 and 8 from www.researchgate.net

However, assays are not always so reliable and examples are readily found of plots of higher dilution factors (higher Cq) having a different gradient to the amplification plots of the lower dilutions (lower Cq). During the exponential phase, the amount of PCR product approximately doubles in each cycle

Amplification plot for Real TimePCR reaction for samples 2, 4, 6 and 8

The amplification plot shows two phases, an exponential phase followed by a non-exponential plateau phase The Real-time qPCR guide: Part 3—troubleshooting uses stylized drawings like the above and real amplification plots to help you identify the factors that could be compromising your results The Amplification Plot screen displays post-run amplification of the samples of each experiment added to your project

Understanding Ct Values In RealTime PCR, 57 OFF. This is because the amplification plots for each dilution of sample are parallel and so the change in threshold affects each amplification plot to the same degree The Amplification Plot screen displays post-run amplification of the samples of each experiment added to your project

Understanding Ct Values In RealTime PCR, 57 OFF. Three plots are available: ΔRn vs Cycle—ΔRn is the magnitude of normalized fluorescence signal generated by the reporter at each cycle during the PCR amplification (ΔRn = Rn - baseline) The C t value increases with a decreasing amount of template