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• W1554018051 abstract "The polymerase chain reaction (PCR) is a primer-mediated, enzymatic amplification of specific genomic DNA or cDNA sequences. RNA-based assays for transcript quantification are arranged vertically, moving down from highest sensitivity to lowest. The sensitivity index ranking is based on three factors: intrinsic difficulty of the technique, reproducibility, and current average level of sensitivity. The final mass of the PCR product(s) is dependent on the efficiency of the reverse transcriptase and the efficiency of the thermostable enzyme(s) used to support PCR, even if said actions occur in the same tube and by the action of the same enzyme. Thus any reliable quantification method must factor in the relative efficiencies of both enzyme-mediated reactions because the overall efficiency of a technique is the product of the efficiencies of the individual steps involved. In “real-time” PCR, which is a fluorescence-based approach, PCR products are quantified as they accumulate, as opposed to performing a fixed number of cycles and then assessing the amount and variety of products by electrophoresis. There are two ways to approach data analysis in real-time PCR applications. The first is the relative quantification method, whereby the amplification of an experimental template is compared to that of a control sample. The other method, known as absolute quantification, is used to calculate the actual copy number of template molecules with statistical confidence; the newest innovation in this area is known as digital PCR. It is very important to include well designed positive, negative, internal, and external controls in all quantitative and semi-quantitative assays. Manuscripts submitted for publication should include information recommended by the MIQE guidelines so as to enable experimental reproducibility." @default.
• W1554018051 created "2016-06-24" @default.
• W1554018051 creator A5003710396 @default.
• W1554018051 date "2010-01-01" @default.
• W1554018051 modified "2023-09-26" @default.
• W1554018051 title "Quantitative PCR Techniques" @default.
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• W1554018051 doi "https://doi.org/10.1016/b978-0-12-374727-3.00019-x" @default.
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