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实时定量聚合酶链反应

Real time quantitative PCR.

作者信息

Heid C A, Stevens J, Livak K J, Williams P M

机构信息

BioAnalytical Technology Department, Genentech, Inc., South San Francisco, California 94080, USA.

出版信息

Genome Res. 1996 Oct;6(10):986-94. doi: 10.1101/gr.6.10.986.

Abstract

We have developed a novel "real time" quantitative PCR method. The method measures PCR product accumulation through a dual-labeled fluorogenic probe (i.e., TaqMan Probe). This method provides very accurate and reproducible quantitation of gene copies. Unlike other quantitative PCR methods, real-time PCR does not require post-PCR sample handling, preventing potential PCR product carry-over contamination and resulting in much faster and higher throughput assays. The real-time PCR method has a very large dynamic range of starting target molecule determination (at least five orders of magnitude). Real-time quantitative PCR is extremely accurate and less labor-intensive than current quantitative PCR methods.

摘要

我们开发了一种新型的“实时”定量PCR方法。该方法通过双标记荧光探针(即TaqMan探针)来测量PCR产物的积累。此方法能对基因拷贝进行非常准确且可重复的定量。与其他定量PCR方法不同,实时PCR不需要PCR后对样品进行处理,避免了潜在的PCR产物污染,从而实现更快且通量更高的检测。实时PCR方法在起始靶分子测定方面具有非常大的动态范围(至少五个数量级)。实时定量PCR极其准确,且比当前的定量PCR方法所需人力更少。

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