44] Chromatography of nucleic acid components on Dowex 1 columns by the use of an automatic recording system

Abstract

Publisher Summary Chromatography using the Dowex 1 column plays an important part in the analysis or preparative method of the study of nucleic acid. Chromatography that uses liquid for migration phase, generally, takes a comparatively long time and it is rather troublesome to determine each fraction collected by the fraction collector. A method of automatic nucleic acid analysis, which separates and analyzes nitrogenous bases, nucleosides, and nucleotides using one Dowex 1-X8-resin column, with an eluting solution of acetate buffer at pH 4.4 within 24 hours, with a linear gradient system is reported. Many instruments determine the absorbency of only one wavelength selecting 2537 A with a filter, and with a mercury lamp as a light source. Before using chromatography, the column temperature is adjusted to 35°, and the column is equilibrated by eluting about 300 ml of 0.1 N acetate buffers at pH 4.4. When the chromatography is finished, the column used is washed by the elution of 3 M acetate buffer (pH 4.4). If no peak is eluted further, the column may be used for new chromatography after the equilibration by 0.1 N acetate buffer pH 4.4. It is desirable to use two columns alternatively for a recording system.