The amount of DNA or cDNA in an unknown sample can then be calculated from its C q value.īaseline: The initial concentration of template is low therefore, the fluorescence intensity is too low to be detected and only the background signal is evident.Įxponential: After the target yield has reached the detection threshold, shown as the red threshold line, the course of the reaction can be followed through the exponential phase. By using multiple dilutions of a known amount of standard DNA, a standard curve can be generated of log concentration against C q. The point at which the fluorescence becomes measurable is called the Quantification Cycle (C q) or crossing point. When the DNA is in the log linear phase of amplification, the amount of fluorescence increases above the background. In quantitative PCR, DNA amplification is monitored at each cycle of PCR.
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