Estimation of DNA Base Composition by High Performance Liquid Chromatography of Its Nuclease PI Hydrolysate

Abstract

In order to develop a new method for determination of the G+C content of a DNA, preparations from 26 bacterial strains and salmon sperm were digested to form 5'-deoxyribonucleotide-monophosphates (dNMP) with nuclease P1 and subjected to high performance liquid chromatography (HPLC). Chromatograms indicated excellent separation of the components, and the data analysis suggested sufficient reproducibility and reasonable A/T and G/C ratios over a wide range (39 to 72mol%) of G+C values.