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The Dot-Blot Technique

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Techniques in Molecular Biology

Abstract

The techniques of Southern and Northern blotting have become widely known and are extensively used in the study of molecular genetics. Briefly, DNA is fragmented by incubation with restriction endonucleases and the fragments are then separated by electrophoresis. The double-stranded DNA in the gel is then denatured and transferred to a filter, usually nitrocellulose. This filter is incubated with a 32P-labelled DNA fragment which has a base sequence complementary to the DNA (or RNA) of interest. This fragment will hybridise (‘stick’) to complementary DNA sequences, while unbound DNA may be rinsed off. The position of hybridisation is subsequently detected by autoradiography. The technique is called Southern blotting in the case of DNA/DNA hybrids and Northern blotting when RNA is subjected to electrophoresis followed by formation of RNA/DNA hybrids.

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Abbreviations

DBM:

Diazobenzyloxymethyl

DPT:

Diazophenylthioether

DEAE:

Diethylaminoethyl

Further Reading

  • Barinaga, M., Franco, R., Meinkoth, J., Ong, E. and Wahl, G.M. (1981) Methods for the Transfer of DNA, RNA and Protein to Nitrocellulose and Diazotized Paper Solid Supports. A Schleicher and Scheull Manual

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  • Bio-Rad Laboratories Manual (1983) Bio-Dot Microfiltration Apparatus Instruction Manual

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  • Bresser, J. and Gillespie, D. (1983) Quantitative Binding of Covalently Closed Circular DNA to Nitrocellulose in Nal, Anal. Biochem., 129, 357–364

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Authors
  1. Paul D. van Helden
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  2. C. Louise Olliver
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Editor information

Editors and Affiliations

  1. Division of Biological and Environmental Sciences, The Hatfield Polytechnic, PO Box 109, College Lane, Hatfield, Hertfordshire, AL10 9AB, England

    John M. Walker (Principal Lecturer in Biochemistry) (Principal Lecturer in Biochemistry)

  2. Department of Infectious Diseases, Section of Bacteriology, University of Utrecht, PO Box 80.171, 3508 TD, Utrecht, The Netherlands

    Wim Gaastra (Associate Professor of Microbiology) (Associate Professor of Microbiology)

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© 1987 John M. Walker and Wim Gaastra

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van Helden, P.D., Olliver, C.L. (1987). The Dot-Blot Technique. In: Walker, J.M., Gaastra, W. (eds) Techniques in Molecular Biology. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-9799-5_10

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  • DOI: https://doi.org/10.1007/978-1-4615-9799-5_10

  • Publisher Name: Springer, Boston, MA

  • Print ISBN: 978-0-7099-3673-2

  • Online ISBN: 978-1-4615-9799-5

  • eBook Packages: Springer Book Archive

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