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Quantitation of mRNA by the polymerase chain reaction.
Open accessAbstract
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.
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Cited by
A new mathematical model for relative quantification in real-time RT-PCR
Pfaffl, M. W. (2001) · Nucleic Acids Research · doi:10.1093/nar/29.9.e45 · Cited by 34,610
Positional cloning of the mouse obese gene and its human homologue
Zhang, Y., Proenca, R., Maffei, M., Barone, M., Leopold, L., & Friedman, J. M. (1994) · Nature · doi:10.1038/372425a0 · Cited by 13,419
Alzheimer-type neuropathology in transgenic mice overexpressing V717F β-amyloid precursor protein
Games, D., Adams, D., Alessandrini, R., Barbour, R., Borthelette, P., Blackwell, C., Carr, T., Clemens, J., Donaldson, T., Gillespie, F., et al. (1995) · Nature · doi:10.1038/373523a0 · Cited by 2,516
Kern, P. A., Saghizadeh, M., Ong, J. M., Bosch, R. J., Deem, R., & Simsolo, R. B. (1995) · Journal of Clinical Investigation · doi:10.1172/jci117899 · Cited by 1,305
Twenty-Five Years of Quantitative PCR for Gene Expression Analysis
VanGuilder, H. D., Vrana, K. E., & Freeman, W. M. (2008) · BioTechniques · doi:10.2144/000112776 · Cited by 1,246
Metadata
- Authors
- Wang, A. M., Doyle, M. V., & Mark, D. F.
- Venue
- Proceedings of the National Academy of Sciences
- Published
- 1989-12
- Volume
- 86
- Issue
- 24
- Pages
- 9717-9721
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- OpenAlex
- W2149897099
- Cited by
- 1,625
- References
- 0
- FWCI
- 24.55
- Type
- journal article
- Language
- en
- Open access
- green
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- CoChat. (2026). Citation check: Quantitation of mRNA by the polymerase chain reaction. https://app.cochat.ai/papers/10.1073/pnas.86.24.9717
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- Wang, A. M., Doyle, M. V., & Mark, D. F. (1989). Quantitation of mRNA by the polymerase chain reaction. Proceedings of the National Academy of Sciences. https://doi.org/10.1073/pnas.86.24.9717
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