An improved direct RNA sequence method; its application to Vicia faba 5.8S ribosomal RNA.
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.
D001483:Base Sequence; D003636:DEAE-Cellulose; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D007887:Fabaceae / Q000235:genetics*; D006801:Humans; D008722:Methods; D010761:Phosphorus Radioisotopes; D010946:Plants, Medicinal; D012329:RNA, Bacterial / Q000235:genetics; D012335:RNA, Ribosomal / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics; D013045:Species Specificity
D010761:Phosphorus Radioisotopes; D012329:RNA, Bacterial; D012335:RNA, Ribosomal; D003636:DEAE-Cellulose