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Nucleic Acids Research, 1975, Vol. 2, No. 10 1911-1930
© 1975

Articles

Hybridization of labeled RNA to DNA in agarose gels

Thomas M. Shinnick*, Elsebet Lund+, Oliver Smithies and Frederick R. Blattner

Laboratory of Genetics, University of Wisconsin Madison, WI, USA *Department of Biology, Massachusetts Institute of Technology Cambridge, MA, USA +Department of Physiological Chemistry, University of Wisconsin Madison, WI, USA

Received September 4, 1975. Specific DNA restriction endonuclease fragments can be identified after electrophoresis in agarose gels by hybridization in the gel (in situ) to radioactive homologous RNA. RNA-DNA hybrids are detected by autoradiography of the gel. Comparison of band patterns of the autoradiogram and the ethidium bromide stained gel allows the identification of the DNA fragment which is complementary to the RNA probe. The technique is rapid, easy and inexpensive. It is sensitive enough to detect individual genes in a mixture of fragments produced by restriction enzyme digestion of complex cellular DNA.

We have used this technique to determine which of the Hin III and Eco Rl fragments of ø80d3 ilv+su+7 and E. coli DNAs contain the 5S, 16S and 23S ribosomal RNA (rRNA) genes of E. coli.


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