Jun 18, 2015 · Let’s start by considering the classical, “normal” qPCR amplification curve shape as shown in Figure 1. As generally plotted, this is a sigmoidal shape with what we’ll refer to as three visually distinct apparent phases or regions.

During RT-qPCR, the accumulated target DNA (expressed as fluorescence) is detected and measured as the reaction progresses, and this information is plotted on a qPCR chart. The qPCR detection methods use a fluorescent signal to measure the amount of DNA in a sample.

To understand how real-time PCR works, we illustrate a qPCR analysis using a typical amplification plot (Figure 1). In this plot, the number of PCR cycles is shown on the x-axis, and the fluorescence from the amplification reaction, which is proportional to the amount of amplified product in the tube, is shown on the y-axis.

Quantitative PCR (qPCR) is one of the foremost methods for detection and quantification of nucleic acid targets. The technique is also known as real-time qPCR and is not to be confused with RT-qPCR (Reverse Transcription-qPCR).

Gene expression analysis by real-time qPCR has been a key enabler of a routine and robust approach for measuring gene expression in genes of interest, as well as monitoring biomarkers. This section will provide the key features of qPCR data analysis and describe examples of common methods to analyze data from a qPCR assay.

Real-time PCR, also called quantitative PCR or qPCR, can provide a simple and elegant method for determining the amount of a target sequence or gene that is present in a sample. Its very simplicity can sometimes lead to problems by overlooking some of …

Oct 9, 2009 · Incorporation of the recommendations from the previous articles into a standard operating procedure will go some way towards generating amplification plots that are not just aesthetically pleasing but are the results of efficient qPCR assays and accurately relate to the original nucleic acid sample.

May 5, 2011 · When something looks off, it’s a red flag to start digging into potential causes and possible corrective actions. The characteristics of the amplification plot can offer information for troubleshooting.

Real-time PCR (also called quantitative PCR or shorter qPCR) can be used to detect the amount of a gene that is present in a sample. The CT (cycle threshold) value of a given gene represents the PCR amplification cycle at which the gene is detected above a certain threshold.

Plots in one or two dimensions are conveniently visualized by human eyes. Plots in three dimensions may also be possible with appropriate tools, but higher dimensional plots are significantly harder to visualize.