Article

Transgenic Research

, Volume 2, Issue 2, pp 115-120

First online:

The use of the nonradioactive digoxigenin chemiluminescent technology for plant genomic Southern blot hybridization: A comparison with radioactivity

  • Gabriele Neuhaus-UrlAffiliated withInstitute for Plant Sciences, Swiss Federal Institute of Technology Zürich
  • , Gunther NeuhausAffiliated withInstitute for Plant Sciences, Swiss Federal Institute of Technology Zürich

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Abstract

A nonradioactive labelling and detection method for plant genomic DNA analysis is compared to the radioactive method. The radioisotopes are replaced by a nucleotide, digoxigenin-11-dUTP, and the signal detection is accomplished by the enzymatic reaction of alkaline phosphatase, conjugated to anti-digoxigenin antibodies, with the chemiluminescent substrate AMPPD (3-(2′-spiroadamantane)-4-methoxy-4(3″ phosphorytoxy) phenyl-1, 2-dioxetane). The sensitivity of the radioactive and nonradioactive methods are directly compared using identical Southern blots subjected to the radioactive and nonradioactive detection. The advantages of this nonradioactive method are discussed.

Keywords

chemiluminescence digoxigenin nonradioactive plant DNA Southern blot hybridization