Rapid group separations of nucleotides and related compounds on silica columns |
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Authors: | Clinton D Lothrop Mayo Uziel |
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Institution: | 1. University of Tennessee—Oak Ridge Graduate School of Biomedical Sciences, Oak Ridge, Tennessee 37830 USA;2. the Biology Division, Oak Ridge National Laboratory, Oak Ridge, Tennessee 37830 USA |
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Abstract: | Nucleosides, bases, and nucleotides can be separated from one another rapidly (10–15 min) on 1-ml silica cartridges. Samples adjusted to 4 mm ammonium borate, 90% acetonitrile are loaded onto 1-ml columns equilibrated with the same solvent. Bases do not absorb to the silica under these conditions. Nucleosides are eluted with 16 ml of 0.5 m acetic acid in 90% acetonitrile. Nucleotides are then eluted with water. The 1-ml silica columns have performed well with samples up to 10 ml in volume. We have found the procedure to be quantitative and the gels to have high capacity (61 μmol Cyd/ml silica). Acid extracts from a large number of cells (108) have been processed on a single cartridge. |
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Keywords: | Address reprint requests to this author |
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