Flow diagram outlining the general procedure for RNA detection by northern blotting.
The northern blot, or RNA blot,[1] is a technique used in molecular biology research to study gene expression by detection of RNA (or isolated mRNA) in a sample.[2][3]
With northern blotting it is possible to observe cellular control over structure and function by determining the particular gene expression rates during differentiation and morphogenesis, as well as in abnormal or diseased conditions.[4] Northern blotting involves the use of electrophoresis to separate RNA samples by size, and detection with a hybridization probe complementary to part of or the entire target sequence. Strictly speaking, the term 'northern blot' refers specifically to the capillary transfer of RNA from the electrophoresis gel to the blotting membrane. However, the entire process is commonly referred to as northern blotting.[5] The northern blot technique was developed in 1977 by James Alwine, David Kemp, and George Stark at Stanford University.[6] Northern blotting takes its name from its similarity to the first blotting technique, the Southern blot, named for biologist Edwin Southern.[2] The major difference is that RNA, rather than DNA, is analyzed in the northern blot.[7]
^Gilbert, S. F. (2000) Developmental Biology, 6th Ed. Sunderland MA, Sinauer Associates.
^ abAlberts, B., Johnson, A., Lewis, J. Raff, M., Roberts, K., Walter, P. 2008. Molecular Biology of the Cell, 5th ed. Garland Science, Taylor & Francis Group, NY, pp 538–539.
^Kevil, C. G., Walsh, L., Laroux, F. S., Kalogeris, T., Grisham, M. B., Alexander, J. S. (1997) An Improved, Rapid Northern Protocol. Biochem. and Biophys. Research Comm. 238:277–279.
^Schlamp, K.; Weinmann, A.; Krupp, M.; Maass, T.; Galle, P. R.; Teufel, A. (2008). "BlotBase: A northern blot database". Gene. 427 (1–2): 47–50. doi:10.1016/j.gene.2008.08.026. PMID 18838116.
^Trayhurn, P. (1996) Northern Blotting. Pro. Nutrition Soc. 55:583–589.
^Alwine JC, Kemp DJ, Stark GR (1977). "Method for detection of specific RNAs in agarose gels by transfer to diazobenzyloxymethyl-paper and hybridization with DNA probes". Proc. Natl. Acad. Sci. U.S.A. 74 (12): 5350–4. Bibcode:1977PNAS...74.5350A. doi:10.1073/pnas.74.12.5350. PMC 431715. PMID 414220.
^Bor, Y.C.; Swartz, J.; Li, Y.; Coyle, J.; Rekosh, D.; Hammarskjold, Marie-Louise (2006). "Northern Blot analysis of mRNA from mammalian polyribosomes". Nature Protocols. doi:10.1038/nprot.2006.216.
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many methods, such as generating hybridization probes for Southern or northernblot hybridization. PCR supplies these techniques with large amounts of pure...
fluorescent protein vector Enzyme assay Protein purification Western blotNorthernblot Southern blot Restriction enzyme Polymerase chain reaction Two-hybrid screening...
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