Rapid and simple comparison of messenger RNA levels using real-time PCR
Rapid and simple comparison of messenger RNA levels using real-time PCR
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DOI：
10.1251/bpo114
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发表时间：
2006-01-25
影响因子：
6.4
通讯作者：
Pouliot, M
中科院分区：
文献类型：
--
作者：
Dussault, AA;Pouliot, M
实时聚合酶链反应（PCR）构成了对传统终点PCR的显著改进，因为它允许实时定量核酸模板的起始量。然而，定量需要通过大量的内部控制和标准曲线进行验证。在本文中，我们描述了一个简单的协议，使用实时PCR比较不同的实验条件之间的感兴趣的基因的mRNA水平。比较实时PCR可以是相对低成本的方法，并且不需要序列特异性荧光报告基因。此外，可以在单次运行中评估来自一组实验的几个基因。因此，除了提供感兴趣的基因表达的比较概况之外，该方法还可以提供关于不同mRNA种类的相对丰度的信息。
Real-time polymerase chain reaction (PCR) constitutes a significant improvement over traditional end-point PCR, as it allows the quantification of starting amounts of nucleic acid templates, in real-time. However, quantification requires validation through numerous internal controls and standard curves. We describe in this paper a simple protocol which uses real-time PCR to compare mRNA levels of a gene of interest between different experimental conditions. Comparative real-time PCR can be a relatively low-cost method and does not require sequence-specific fluorescent reporters. Moreover, several genes from a set of experiments can be assessed in a single run. Thus, in addition to providing a comparative profile for the expression of a gene of interest, this method can also provide information regarding the relative abundance of different mRNA species.