Micropellicular stationary phases for high-performance liquid chromatography of double-stranded DNA.

The central role of nucleic acids in biosciences has effectuated the rapid development of numerous techniques for their isolation, separation, characterization and quantitation. Advances in high-performance liquid chromatography, particularly the introduction of novel microparticulate sorbents, have greatly promoted the separation and quantitation of nucleic acids. Because of their favorable mass transfer properties, micropellicular packing materials are advantageous for fast and high-resolution separations of double-stranded (ds) DNA molecules. With micropellicular packings, anion-exchange and ion-pair reversed-phase chromatography are the most popular chromatographic separation modes for dsDNA. The effective separation mechanisms in both chromatographic techniques are preferably described by nonstoichiometric models, that are founded on a better physicochemical background than traditional stoichiometric models. Column efficiency, retention characteristics, and size or sequence dependency of retention of dsDNA are greatly influenced by the chosen operational variables in both chromatographic modes. The applicability of HPLC with micropellicular stationary phases nucleic acids research includes preparative DNA fractionation, DNA restriction mapping, analysis of polymerase chain reaction products and purification of plasmid DNA.