Separation of DNA fragments by isocratic HPLC

Isocratic high-performance liquid chromatography (HPLC) was used to separate single stranded DNA fragments. Different concentrations of acetonitrile in 0.1 triethylammonium acetate were used to separate DNA fragments differing in length by 100 base pairs. Various single stranded DNA molecules ranging from 100 to 1500 base pairs were used for this study. Optimum isocratic HPLC conditions for this separation were determined on a Hamilton PRP Polystyrene column. The best separation was obtained when the concentration of acetonitrile was 25-35%. With this concentration eluent small DNA fragments were separated from one another, but the large DNA fragments in the sample were retained on the column.