Quantitation of mRNA by the polymerase chain reaction.

AM Wang, MV Doyle, DF Mark - Proceedings of the …, 1989 - National Acad Sciences
AM Wang, MV Doyle, DF Mark
Proceedings of the National Academy of Sciences, 1989National Acad Sciences
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 …
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.
National Acad Sciences