A new mathematical model for relative quantification in real-time RT-PCR

We focused on the relative quantification of target gene transcripts in comparison to a reference gene transcript

M. W. Pfaffl

2002

Scholarcy highlights

  • Reverse transcription followed by the polymerase chain reaction is the technique of choice to analyse mRNA expression derived from various sources
  • To investigate the physiological changes in gene expression, the relative expression ratio is adequate for the most purposes. An absolute quantification, based either on an internal or an external calibration curve
  • Specificity of RT–PCR products was documented with high resolution gel electrophoresis and resulted in a single product with the desired length
  • Real-time PCR efficiencies were calculated from the given slopes in LightCycler software
  • We focused on the relative quantification of target gene transcripts in comparison to a reference gene transcript
  • The established mathematical model is presented in order to better understand the mode of analysis in relative quantification in real-time RT
  • polymerase chain reaction in general and the minimal error rate of the model allows for an accurate determination of the relative expression ratio

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