Summary
A novel approach of competitive PCR for quantifying mRNAs is described where competitors are constructed by deleting an internal sequence from target reverse transcribed mRNAs. The two sets of product generated (control and target) are easily discriminated by size and quantified for the sequence of interest and a housekeeping gene, β-actin,used as internal mRNA control.
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Becker-Andre, M. & Halbrock, K. (1989) Nucl. Acids Res. 17, 9437–9446.
Chelly, J., Kaplan, J. C., Maire, P., Gautron, S. & Kahn, A. (1988). Nature 333, 858–860.
Chomczinski, P, and Sacchi, N. (1987). Anal. Biochem. 162, 159–162.
Diviacco, S., Norio, P., Zentilin, L., Menzo, S., Clementi, M., Biamonti, G., Riva, S., Falaschi, A. and Giacca, M (1992). Gene 122, 313–320.
Foley, K.P., Leonard, M.W., and Engel, J.D. (1993).Trends Genet. 9, 380–385.
Frohman, M. A., Dush, M.K., and Martin, G.R. (1988). Proc. Natl. Acad. Sci. USA 85, 8998–9002.
Gilliland, G., Perrin, S., Blanchard, K. and Bunn, H.F. (1990a). Proc. Natl. Acad. Sci. USA 87, 2725–2729.
Gilliland, G., Perrin, S., and Bunn, H.F. (1990b). In: PCR Protocols, M.A. Innis et al., eds. pp. 60–69, San Diego, California, Academic Press,Inc.
Guillem, J.G., Levy, M.F., Hsieh, L.L., Johnson, M.D., LoGerfo, P., Forde, K.A., and Weinstein, I.B. (1990). Mol. Carcinog. 3, 68–74.
Horikoshi, T., Danenberg, K.D., Stadlbauer, T.H.W., Volkenandt, M., Shea, L.C.C., Aigner, K., Gustavsson, B., Leichman, L., Frösing, R., Ray, M., Gibson, N.W., Spears, C.P., and Danenberg, P.V. (1992). Cancer Res. 52, 108–116.
Nakajima-Iijima, S., Hamada, H., Reddy, P., and Kakunaga, T. (1985).Proc. Natl. Acad. Sci. USA 82, 6133–6137.
Siebert, P.D. and Larrick, J.W. (1993). Biotechniques 14, 244–249.
Van Straten, F., Müller, R., Curran, T., VanBeveren, C., and Verma, I.M. (1983). Proc. Natl. Acad. Sci. USA 80, 3183–3187.
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Del Giacco, L., Doneda, L., Grammatico, P. et al. Rapid construction of competitive templates for quantitative PCR of reverse-transcribed mRNAs.. Biotechnol Tech 9, 879–884 (1995). https://doi.org/10.1007/BF00158540
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DOI: https://doi.org/10.1007/BF00158540