Northern Blotting Analysis
Purchase on Springer.com
$49.95 / €39.95 / £34.95*
* Final gross prices may vary according to local VAT.
Northern blotting analysis is a classical method for analysis of the size and steady-state level of a specific RNA in a complex sample. In short, the RNA is size-fractionated by gel electrophoresis and transferred by blotting onto a membrane to which the RNA is covalently bound. Then, the membrane is analysed by hybridization to one or more specific probes that are labelled for subsequent detection. Northern blotting is relatively simple to perform, inexpensive, and not plagued by artefacts. Recent developments of hybridization membranes and buffers have resulted in increased sensitivity closing the gap to the more laborious nuclease protection experiments.
- Alwine, J. C., Kemp, D. J., Stark, G. R. (1977) Method for detection of specific RNAs in agarose gels by transfer to diazobenzyloxymethyl-paper and hybridization with DNA probes. Proc Natl Acad Sci USA 74, 5350–5354. CrossRef
- Alwine, J. C., Kemp, D. J., Parker, B. A., Reiser, J., Renart, J., Stark, G. R., Wahl, G. M. (1979) Detection of specific RNAs or specific fragments of DNA by fractionation in gels and transfer to diazobenzyloxymethyl paper. Methods Enzymol 68, 220–242. CrossRef
- Southern, E. M. (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98, 503–517. CrossRef
- Lamond, A. I., Sproat, B. S. (1994) Isolation and characterization of Ribonucleoprotein complexes, in (Higgins, S. J. and Hames, B. D., eds.) RNA Processing. A Practical Approach. IRL Press, Oxford, Vol. 1, pp. 103–140.
- Sambrook, J., Fritsch, E. F., Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
- Darling, D. C., Brickell, P. M. (1994) Nucleic Acid Blotting. The Basics.. IRL Press, Oxford.
- Farrell, R. E.,Jr. (1993) RNA Methodologies. A Laboratory Guide for Isolation and Characterization. Academic, San Diego, CA.
- Chomczynski, P., Sacchi, N. (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162, 156–159. CrossRef
- Aviv, H., Leder, P. (1972) Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci USA 69, 1408–1412. CrossRef
- Lehrach, H., Diamond, D., Wozney, J. M., Boedtker, H. (1977) RNA molecular weight determinations by gel electrophoresis under denaturing conditions, a critical reexamination. Biochemistry 16, 4743–4751. CrossRef
- McMaster, G. K., Carmichael, G. G. (1977) Analysis of single- and double-stranded nucleic acids on polyacrylamide and agarose gels by using glyoxal and acridine orange. Proc Natl Acad Sci USA 74, 4835–4838. CrossRef
- Reijnders, L., Sloof, P., Sival, J., Borst, P. (1973) Gel electrophoresis of RNA under denaturing conditions. Biochim Biophys Acta 324, 320–333.
- Chomczynski, P., Mackey, K. (1994) One-hour downward capillary blotting of RNA at neutral pH. Anal Biochem 221, 303–305. CrossRef
- Bittner, M., Kupferer, P., Morris, C. F. (1980) Electrophoretic transfer of proteins and nucleic acids from slab gels to diazobenzyloxymethyl cellulose or nitrocellulose sheets. Anal Biochem 102, 459–471. CrossRef
- Herrin, D. L., Schmidt, G. W. (1988) Rapid, reversible staining of northern blots prior to hybridization. Biotechniques 6, 196–200.
- Church, G. M., Gilbert, W. (1984) Genomic sequencing. Proc Natl Acad Sci USA 81, 1991–1995. CrossRef
- Saito, I., Sugiyama, H., Furukawa, N., Matsuura, T.. (1981) Photoreaction of thymidine with primary amines. Application to specific modification of DNA. Nucleic Acids Symp Ser 10, 61–64.
- Feinberg, A. P., Vogelstein, B.. (1983) A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132, 6–13. CrossRef
- Casey, J., Davidson, N. (1977) Rates of formation and thermal stabilities of RNA:DNA and DNA:DNA duplexes at high concentrations of formamide. Nucleic Acids Res 4, 1539–1552. CrossRef
- Bonner, J., Kung, G., Bekhor, I. (1967) A method for the hybridization of nucleic acid molecules at low temperature. Biochemistry 6, 3650–3653. CrossRef
- Northern Blotting Analysis
- Book Title
- Book Subtitle
- Methods and Protocols
- pp 87-105
- Print ISBN
- Online ISBN
- Series Title
- Methods in Molecular Biology
- Series Volume
- Series Subtitle
- Methods and Protocols
- Series ISSN
- Humana Press
- Copyright Holder
- Springer Science+Business Media, LLC
- Additional Links
- Gel electrophoresis
- northern blotting
- hybridization analysis
- Industry Sectors
- Henrik Nielsen (ID1)
- Editor Affiliations
- ID1. The Panum Institute, Dept. of Cellular & Molecular Medicine, University of Copenhagen
- Author Affiliations
- 1. The Bartholin Institute, Copenhagen University Hospital, Copenhagen, Denmark
- 2. Department of Cellular and Molecular Medicine, The Panum Institute, University of Copenhagen, Copenhagen, Denmark
To view the rest of this content please follow the download PDF link above.