Transcription of Relative quantification
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Relative quantification
Relative quantificationMichael W. Pfaffl in: real - time PCR. Published by International University Line (Editor: T. Dorak), p IntroductionReverse transcription (RT) followed by a polymerase chain reaction (PCR)represents the most powerful technology to amplify and detect traceamounts of mRNA (Heid et al., 1996; Lockey, 1998). To quantify these lowabundant expressed genes in any biological matrix the real - time quantita-tive RT-PCR (qRT-PCR) is the method of choice. real - time qRT-PCR hasadvantages compared with conventionally performed semi-quantitativeend point RT-PCR, because of its high sensitivity, high specificity, goodreproducibility, and wide dynamic quantification range (Higuchi et al.,1993; Gibson et al., 1996; Orland et al., 1998; Freeman et al.)
Two general types of quantification strategies can be performed in qRT-PCR. The levels of expressed genes may be measured by an ‘absolute’ quantification or by a relative or comparative real-time qRT-PCR (Pfaffl, 2004). The ‘absolute’ quantification approach relates the PCR signal to
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