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Nucleic Acids Research, 1980, Vol. 8, No. 6 1259-1272
© 1980

Articles

An improved direct RNA sequence method; its application to Vida faba 5.8S ribosomal RNA

Y. Tanaka, T.A. Dyer* and G.G. Brownlee

MRC Laboratory of Molecular Biology Hills Road, Cambridge CB2 2QH *ARC Plant Breeding Institute Maris Lane, Trumpington, Cambridge CB2 2LQ, UK

Received January 31, 1980. We have developed a direct read-off sequencing procedure, based on the method of Stanley and Vassilenko using E. coli 5S ribosomal RNA as a model compound. Radioactive bands were transferred from an acrylamide gel fractionation in the first dimension onto a DEAE-cellulose thin layer plate. After in situ enzymatic digestion with RNase T2, mononucleoside 3',5'-di-phosphates were separated in the second dimension by electrophoresis at pH 2.3. Using this two-dimensional procedure the entire sequence of 163 residues of the previously unknown Vicia faba (broad bean) 5.8S ribosomal RNA was deduced.


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