ABSOLUTE QUANTIFICATION OF TARGET DNA - A SIMPLE COMPETITIVE PCR FOR EFFICIENT ANALYSIS OF MULTIPLE SAMPLES
ABSOLUTE QUANTIFICATION OF TARGET DNA - A SIMPLE COMPETITIVE PCR FOR EFFICIENT ANALYSIS OF MULTIPLE SAMPLES
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DOI：
10.1093/nar/21.8.2017
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发表时间：
1993-04-25
影响因子：
14.9
通讯作者：
GOUSTIN, AS
中科院分区：
文献类型：
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作者：
ZACHAR, V;THOMAS, RA;GOUSTIN, AS
PCR是一种灵敏度极高的检测微量靶DNA的方法。然而，DNA扩增的指数性质易于使实验数据负担显著的标准误差，这是由于扩增效率的管与管之间的固有变化。因此，通过PCR对DNA或RNA模板进行可靠的定量需要引入内标。通常，在分析样品的一系列重复中，将限定范围的此类标准品与恒定未知量的野生型模板一起共扩增（1，2），或者将两种模板进行终点稀释（3）。显然，每种方法都是基于多个PCR反应，这些反应将与每个被分析的样品同时进行。然而，当要分析大量样品时，这样的程序可能变得麻烦。
PCR is an exquisitely sensitive method for detection of minute amounts of target DNA. The exponential nature of DNA amplification, however, is prone to burden the experimental data with significant standard error due to the tube-to-tube inherent variations in amplification efficiency. Therefore the reliable quantitation of DNA or RNA templates by PCR necessitates introduction of an internal standard. Typically, in a series of replicates of analyzed sample the defined range of such standards is coamplified together with constant unknown amount of wildtype template (1, 2) or alternatively both templates are subjected to endpoint dilution (3). Obviously, each of the methods is based on multiple PCR reactions which are to beperformed simultaneously with each of the analyzed samples. Such procedure, nevertheless, may become cumbersome when large numbers of samples are to be analyzed.