Combined pH gradient and anion-exchange high-performance liquid chromatographic separation of oligodeoxyribonucleotides.
A novel method of elution using a pH gradient to separate small thymidine- and guanosine-containing oligonucleotides on a Pharmacia Mono Q HR anion-exchange column is described. The method is based on the alkaline titration of ring protons of the thymine and guanine base moieties and results in excellent separations of di-, tri- and tetranucleotides that either are not resolved in salt gradients near neutral pH or require long elution times when salt gradient elution is used with strongly alkaline eluents.
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Department of Biochemical and Biophysical Sciences, University of Houston, TX 77204-5934.
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D000837:Anion Exchange Resins; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002852:Chromatography, Ion Exchange / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D009838:Oligodeoxyribonucleotides / Q000302:isolation & purification*
D000837:Anion Exchange Resins; D009838:Oligodeoxyribonucleotides