Nonradioactive Probes

  • Peter G. Isaac


When first published in 1975, RFLP procedures used probes labelled with radioisotopes to detect complementary sequences in complex mixtures of restriction fragments (1). During the early 1980s, several groups experimented with nonradioactive probing systems (2, 3). Since then, several commercially available kits have been released that are at least as sensitive as radioisotopes, and which yield results in much shorter times. It is beyond the scope of this section to provide the reader with detailed protocols for the use of nonradioactive probes, but the subject has been reviewed recently (4) and general textbooks are available (5).


Complementary Sequence Detailed Protocol Antibody Conjugate Charge Nylon Membrane Careful Handling 
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  1. 1.
    Southern, E.M.(1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. Journal of Molecular Biology, 98, 503–517.CrossRefGoogle Scholar
  2. 2.
    Leary, J.J., Brigati, D.J. and Ward, D.C. (1983) Rapid and sensitive colorimetric method for visualizing biotin-labelled DNA probes hybridized to DNA or RNA immobilized on nitrocellulose: Bio-blots. Proceedings of the National Academy of Sciences, USA 80, 4045–4049CrossRefGoogle Scholar
  3. 3.
    Ambros P.F., Matzke M.A. and Matzke A.J.M. (1986) Detection of a 17kB unique sequence (T-DNA) in plant chromosomes by in situ hybridization. Chromosoma 94, 11–18CrossRefGoogle Scholar
  4. 4.
    Isaac PG Stacey J, and Clee CM(1995) Nonradioactive probe. Molecular Biotechnology 3, 259–265CrossRefGoogle Scholar
  5. 5.
    Methods in Molecular Biology, vol 28: Protocols for Nucleic Acids analysis by nonradioactive probes (1994) (Isaac PG, ed.). Humana Press, Totowa, New Jersey, USAGoogle Scholar
  6. 6.
    Alderton R.R, Kitau J. and Beck S. (1994) Automated DNA hybridization. Analytical Biochemistry 218, 98–102CrossRefGoogle Scholar
  7. 7.
    Patel A., Martin C, and Gallagher S (1994) New Gel/membrane processor automates nonradioactive detection techniques for blotting targets. Hoefer News 94–1, 1–2Google Scholar

Copyright information

© Chapman & Hall 1998

Authors and Affiliations

  • Peter G. Isaac

There are no affiliations available

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