High-performance affinity chromatography of DNA.

The octadecamer of thymidylic acid, (dT)18, was synthesized with a primary amino group on the 5'-terminal phosphate and this was covalently coupled to 300 A pore macroporous silica. Coupling was performed inside a prepacked column to an activated N-hydroxysuccinimidyl ester silica. The (dT)18-silica column successfully separates mixtures of adenine oligomers differing in length by one nucleotide. The dependence upon salt concentration, temperature and length for elution of oligonucleotides was determined. Methods were also developed to selectively elute such columns using either salt or temperature gradients.