Introduction To Real-Time Quantitative PCR (qPCR - ResearchGate
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Genomic Platform
Institute of research in immunology and cancer, University of Montreal www.genomique.iric.ca 1Information on qPCR results
What does qPCR measure?
If you are measuring gene expression, qPCR will tell you how much of a specific mRNA there is in your samples. You amplify a small region of this mRNA with oligos and a fluorescent probe. The qPCR machine measures the intensity of fluorescence emitted by the probe at each cycle.During the first cycles, there is not enough fluorescence to be detected, but the reaction
rapidly produces more and more amplicons and the fluorescence builds up. A qPCR curve has typically an exponential phase followed by a plateau phase. The Ct measure needs to be taken in the exponential phase, where the curve is linear. Most of the time, a qPCR experiment will give a ³relative H[SUHVVLRQ´, which is a variation of the expression of a gene between two samples.