Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the 2 2 DD C T Method
Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the 2 2 DD C T Method
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通讯作者：
K. Livak;Thomas D. Schmittgen
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作者：
K. Livak;Thomas D. Schmittgen
两种最常用的分析数据的方法，如未处理的对照或样品在时间零点实时定量PCR实验是绝对定量-在时间过程研究中。和相对定量。绝对定量测定-绝对定量应在原位进行，通常通过将PCR信号与标准曲线相关联来确定输入的拷贝数。相对定量将PCR信号作用与另一个转录本拷贝数的PCR信号作用相关联，其中需要确定处理组中靶转录本的绝对值。绝对定量具有样品，例如未处理的对照。2 2 DD C T法是一种结合实时PCR和许多方便的方法来分析文献中出现的基因表达的相对变化，包括来自实时定量PCR实验。这两篇文章的目的是在这个问题上11）。在某些情况下，本文将介绍2 - 2 DD C T法的推导、假设和应用。此外，我们提出的推导和脚本拷贝数和报告的相对变化的应用程序的2 - 2 DD C T的方法，可能是有用的实时，定量PCR数据的分析的两个变化。
of the target gene relative to some reference group The two most commonly used methods to analyze data from such as an untreated control or a sample at time zero real-time, quantitative PCR experiments are absolute quantifica-in a time-course study. tion and relative quantification. Absolute quantification deter-Absolute quantification should be performed in situ- mines the input copy number, usually by relating the PCR signal to a standard curve. Relative quantification relates the PCR signal ations where it is necessary to determine the absolute of the target transcript in a treatment group to that of another transcript copy number. Absolute quantification has sample such as an untreated control. The 2 2 DD C T method is a combined with real-time PCR and numerous re- convenient way to analyze the relative changes in gene expression appeared in the literature including from real-time quantitative PCR experiments. The purpose of this two articles in this issue 11). In some situations, report is to present the derivation, assumptions, and applications it may be unnecessary to determine the absolute tran- of the 2 2 DD C T method. In addition, we present the derivation and script copy number and reporting the relative change applications of two variations of the 2 2 DD C T method that may be useful in the analysis of real-time, quantitative PCR data.