Quantitative real-time RT-PCR data analysis: current concepts and the novel “gene expression's C T difference” formula
JH Schefe, KE Lehmann, IR Buschmann… - Journal of molecular …, 2006 - Springer
JH Schefe, KE Lehmann, IR Buschmann, T Unger, H Funke-Kaiser
Journal of molecular medicine, 2006•SpringerFor quantification of gene-specific mRNA, quantitative real-time RT-PCR has become one of
the most frequently used methods over the last few years. This article focuses on the issue of
real-time PCR data analysis and its mathematical background, offering a general concept for
efficient, fast and precise data analysis superior to the commonly used comparative CT (ΔΔC
T) and the standard curve method, as it considers individual amplification efficiencies for
every PCR. This concept is based on a novel formula for the calculation of relative gene …
the most frequently used methods over the last few years. This article focuses on the issue of
real-time PCR data analysis and its mathematical background, offering a general concept for
efficient, fast and precise data analysis superior to the commonly used comparative CT (ΔΔC
T) and the standard curve method, as it considers individual amplification efficiencies for
every PCR. This concept is based on a novel formula for the calculation of relative gene …
Abstract
For quantification of gene-specific mRNA, quantitative real-time RT-PCR has become one of the most frequently used methods over the last few years. This article focuses on the issue of real-time PCR data analysis and its mathematical background, offering a general concept for efficient, fast and precise data analysis superior to the commonly used comparative C T (ΔΔC T ) and the standard curve method, as it considers individual amplification efficiencies for every PCR. This concept is based on a novel formula for the calculation of relative gene expression ratios, termed GED (Gene Expression’s C T Difference) formula. Prerequisites for this formula, such as real-time PCR kinetics, the concept of PCR efficiency and its determination, are discussed. Additionally, this article offers some technical considerations and information on statistical analysis of real-time PCR data.
Springer