A Two-dimensional Fractionation Procedure for Radioactive Nucleotides

1989 
A method is described for the two-dimensional fractionation of ribonuclease digests of 32P-labelled RNA. High-voltage ionophoresis is used in both dimensions. The first is on cellulose acetate at pH 3·5, the second on DEAE-paper at an acid pH. The method is capable of resolving the di- and tri- and most of the tetra-nucleotides in digests prepared by the action of ribonuclease T1 or pancreatic ribonuclease. It has been applied to the 16 s and 23 s components of ribosomal RNA which show significant quantitative differences, and to sRNA from Escherichia coli and from yeast. A method is described for the determination of the sequence of a nucleotide by partial digestion with spleen phosphodiesterase.
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