Sequence analysis of synthetic oligonucleotides by high-performance liquid anion-exchange chromatography

A simple procedure for the sequential anlysis of small oligonucleotides is reported. The method is based on the simultaneous identification and quantitation of monomers released by venom phosphodiesterase digestion of oligonucleotides using high-performance anion-exchange chromatography on Permaphase AAX at room temperature and by applying isocratic elution conditions. In this way, the correct sequence of five oligomers, e.g., r-ACCUCC, r-CUGUU, r-AGGA, d-ATTACC and d-GGTAAT, could easily be established unambiguously.