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Comparative Study
. 1980 Mar 25;8(6):1259-72.
doi: 10.1093/nar/8.6.1259.

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

Y Tanaka, T A Dyer, G G Brownlee
Free PMC article
Comparative Study

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

Y Tanaka et al. Nucleic Acids Res. .
Free PMC article

Abstract

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'-diphosphates 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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