MICROPELLICULAR STATIONARY PHASES FOR HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY OF DOUBLE-STRANDED DNA

The central role of nucleic acids in biosciences has effectuated the r apid development of numerous techniques for their isolation, separatio n, characterization and quantitation. Advances in high-performance liq uid chromatography, particularly the introduction of novel micropartic ulate sorbents, have greatly promoted the separation and quantitation of nucleic acids. Because of their favorable mass transfer properties, micropellicullar packing materials are advantageous for fast and high -resolution separations of double-stranded (ds) DNA molecules. With mi cropellicular packings, anion-exchange and ion-pair reversed-phase chr omatography are the most popular chromatographic separation modes for dsDNA. The effective separation mechanisms in both chromatographic tec hniques are preferably described by nonstoichiometric models, that are founded on a better physicochemical background than traditional stoic hiometric models. Column efficiency, retention characteristics, and si ze or sequence dependency of retention of dsDNA are greatly influenced by the chosen operational variables in both chromatographic modes. Th e applicability of HPLC with micropellicular stationary phases nucleic acids research includes preparative DNA fractionation, DNA restrictio n mapping, analysis of polymerase chain reaction products and purifica tion of plasmid DNA. (C) 1998 Elsevier Science B.V.