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Nucleic Acids Research, 1974, Vol. 1, No. 8 1059-1068
© 1974

Articles

Deoxynucleotide substitution: a new technique for sequence analysis of RNA

A. Van de Voorde, R. Rogiers, J. Van Herreweghe, H. Van Heuverswyn, G. Volckaert and W. Fiers

Laboratory of Molecular Biology, State University of Ghent Ledeganckstraat 35, 9000 Gent, Belgium

Received July 12, 1974. It is possible to replace in a normal transcription reaction catalyzed by E.coli RNA polymerase one of the four precursors by the corresponding deoxynucleoside triphosphate. These deoxynucleotide-substituted RNA's offer interesting prospects for nucleotide sequence analysis. Indeed by the use of U2-RNase with dG-RNA, or pancreatic RNase with dC-RNA or dU-RNA, base specific cleavage can be obtained at any of the four residues. In this way overlap of at least six residues in length can be obtained for any site in the RNA. The technique offers also great benefit for solving the sequence of the more difficult T1-oligonucleotides. Some examples in the sequence analysis of SV4O DNA-Hind fragments are reported.


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