Determination of DNA base composition by reversed-phase high-performance liquid chromatography |
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| Authors: | Jin Tamaoka Kazuo Komagata |
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| Affiliation: | Institute of Applied Microbiology, University of Tokyo, Bunkyo-ku, Tokyo, 113 Japan |
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| Abstract: | Abstract DNA base composition was determined by reversed-phase high-performance liquid chromatography (HPLC). DNA was hydrolysed into nucleosides with nuclease P1 and bacterial alkaline phosphatase. The mixture of nucleosides was applied to HPLC without any further purification. One determination by chromatography needed 2 μg of hydrolysed nucleosides and took only 8 min. The relative standard error of nucleoside analysis was less than 1%. The system described here gives a direct and precise method for determining DNA base composition. |
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| Keywords: | Enzymatic hydrolysis of DNA nucleoside nuclease P1 bacterial alkaline phosphatase |
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