Abstract
Northern-blot hybridization, also referred to as Northern blotting, is one of several methods developed to detect the presence, to determine the size, and to quantify specific cellular mRNAs. By this method, total RNA or poly(A)+mRNA, prepared from the cells or tissue of interest, is fractionated by size on a denaturing agarose gel. The separated RNAs are transferred to a membrane by capillary action or under a vacuum and hybridized to a labeled probe with a base sequence complementary to all or part of the target mRNA. Analysis of hybridization signals determines whether the gene of interest is expressed in the cells or tissue, as well as the size and relative quantity of the target mRNA, if appropriate markers are used.
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References
Zyskind, J. W. and Bernstein, S.I. (1989) Gel blotting, probe preparation, hybridization and hybrid detection, in Recombinant DNA Laboratory Manual, Academic, San Diego, CA, pp. 118–134
Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York.
Krumlauf, R. (1996) Northern blot analysis, in Methods in Molecular Biology, Volume 58: Basic DNA and RNA Protocols, (Harwood, A. J., ed.), Humana, Totowa, NJ, pp. 113–118.
Trayhurn, P. (1996) Northern blotting. Proc. Nutr. Soc. 55, 583–589.
Greenberg, M. E. and Bender, T. P. (1999) Preparation and analysis of RNA, in Current Protocols in Molecular Biology (Ausubel, F. M., Brent, R., Kingston, R. E., Moore, D. D., Seidman, J. G., Smith, J. A., and Struhl, K., eds.), Wiley, New York, pp. 4.0.1–4.10.11.
Lassegue, B., Alexander, R. W., Nickenig G., Clark M., Murphy T. J., Griendling K. K. (1995) Angiotensin II down-regulates the vascular smooth muscle AT1 receptor by transcriptional and post-transcriptional mechanism: evidence for homologous and heterologous regulation. Mol. Pharmacol. 48, 601–609.
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Gallagher, P.E., Diz, D.I. (2001). Analysis of RNA by Northern-Blot Hybridization. In: Wang, D.H. (eds) Angiotensin Protocols. Methods in Molecular Medicine™, vol 51. Humana Press. https://doi.org/10.1385/1-59259-087-X:205
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DOI: https://doi.org/10.1385/1-59259-087-X:205
Publisher Name: Humana Press
Print ISBN: 978-0-89603-763-2
Online ISBN: 978-1-59259-087-2
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