Non-radioactive detection methods for nucleic acids separated by electrophoresis.
The different non-radioactive labelling and detection methods currently commercially available are compared and evaluated in this review. Minor factors such as electrophoresis and blotting techniques as well as choice of membrane and their impact on results are discussed. Two major labelling moieties, biotin and digoxigenin, and the various labelling methods are discussed in detail. A comparison of my own results and those from the literature favours application of the digoxigenin group as a routine label. Nevertheless, in several cases biotin will also lead to good results and may also serve as a second label. The most important factor within the non-radioactive systems is the detection of the targeted label. Colorimetric and chemiluminescent techniques are compared in terms of sensitivity, flexibility and applicability. Colorimetric detection can produce suitable results, but in most cases the major advantages of chemiluminescent techniques involving alkaline phosphatase and AMPPD or CSPD will make chemiluminescent detection the method of choice. A survey is given on applicability of the basic techniques to several important assay methods involving electrophoresis of nucleic acids. Finally, some examples of application of non-radioactive nucleic acid labelling and detection techniques in plant molecular biology and biomedicine are cited from the literature.
Düring
K
K
Center for Applied Plant Molecular Biology, University of Hamburg, Germany.
D000818:Animals; D004586:Electrophoresis / Q000379:methods*; D006801:Humans; D009696:Nucleic Acids / Q000032:analysis*
D009696:Nucleic Acids