Thermal elution chromatography of nucleic acids on hydroxyapatite

Hydroxyapatite thermal elution chromatography was studied from an empirical standpoint. The dependence of elution temperature on elution buffer concentration was determined for various types of buffer, hydroxyapatite and nucleic acid. The results are analyzed in terms of the proper design and interpretation of thermal elution experiments. The potential for serious artifacts is demonstrated and the means by which they may be avoided is described. Various commercially available hydroxyapatites were tested in conjunction with various aqueous and partially non-aqueous buffer systems. Among the materials tested, potassium phosphate and Bio-Rad HTP were found to constitute the best buffer-hydroxyapatite system for most types of thermal elution study.