Wang A M, Doyle M V, Mark D F
Department of Molecular Biology, Cetus Corporation, Emeryville, CA 94608.
Proc Natl Acad Sci U S A. 1989 Dec;86(24):9717-21. doi: 10.1073/pnas.86.24.9717.
A method for the quantitation of specific mRNA species by the polymerase chain reaction (PCR) has been developed by using a synthetic RNA as an internal standard. The specific target mRNA and the internal standard are coamplified in one reaction in which the same primers are used. The amount of mRNA is then quantitated by extrapolating against the standard curve generated with the internal standard. The synthetic internal standard RNA consists of a linear array of the sequences of upstream primers of multiple target genes followed by the complementary sequences to their downstream primers in the same order. This quantitative PCR method provides a rapid and reliable way to quantify the amount of a specific mRNA in a sample of less than 0.1 ng of total RNA. In addition, the same internal standard RNA is used, with appropriate primer pairs, to quantitate multiple different mRNA species.
通过使用合成RNA作为内标，已开发出一种通过聚合酶链反应（PCR）对特定mRNA种类进行定量的方法。特定的靶mRNA和内标在一个使用相同引物的反应中共同扩增。然后通过根据用内标生成的标准曲线进行外推来定量mRNA的量。合成内标RNA由多个靶基因上游引物序列的线性阵列组成，其后是与其下游引物互补的序列，顺序相同。这种定量PCR方法提供了一种快速可靠的方法，可对总RNA少于0.1 ng的样品中的特定mRNA量进行定量。此外，使用相同的内标RNA和适当的引物对，可对多种不同的mRNA种类进行定量。
