期刊
JOURNAL OF MOLECULAR MEDICINE-JMM
卷 84, 期 11, 页码 901-910出版社
SPRINGER HEIDELBERG
DOI: 10.1007/s00109-006-0097-6
关键词
quantification; expression ratio; PCR efficiency; TaqMan; SYBR Green
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) (Delta Delta C (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.
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