Preparative two-dimensional polyacrylamide gel electrophoresis of 32 P-labeled RNA |
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Authors: | R de Wachter W Fiers |
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Affiliation: | Laboratory of Physiological Chemistry and Laboratory of Molecular Biology, State University of Ghent, Ghent, Belgium |
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Abstract: | Complex mixtures of RNA molecules may be separated by two-dimensional electrophoresis on polyacrylamide gel slabs. The first dimension of the separation is carried out on acid gels in the presence of a high urea concentration, the second on more concentrated gels buffered at pH 8. The method has been applied to the complete separation of RNA fractions obtained after a preliminary gel electrophoresis of partial enzymic digests of 32P-labeled bacteriophage RNA. Another application is the fractionation of partial digests as obtained in sequence determination of RNA molecules. Spots are detected by autoradiography and extracted by a simple micro procedure which yields the material in a concentrated form suitable for sequence analysis by fingerprinting. |
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