Optimal extraction conditions for high-performance liquid chromatographic determination of nucleotides in yeast |
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Authors: | Z Olempska-Beer E B Freese |
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Affiliation: | Laboratory of Molecular Biology, National Institute of Neurological and Communicative Disorders and Stroke, National Institutes of Health, Building 36, Room 3D02, Bethesda, Maryland 20205 USA |
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Abstract: | Different extraction methods of nucleotides from the yeast Saccharomyces cerevisiae were compared. A new extraction solution--formic acid saturated with 1-butanol--was found to be more effective than the commonly used solutions of trichloroacetic acid, perchloric acid, or formic acid alone. Using this solution the optimal extraction conditions were established. Nucleotide recovery was evaluated by adding standard nucleotides to the extraction medium and carrying them together with the cells through the whole extraction procedure. Nucleotides were separated and quantitated by high-performance liquid chromatography on an anion-exchange column. |
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Keywords: | nucleotides extraction recovery evaluation yeast formic acid HPLC |
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