Size-dependent chromatographic separation of nucleic acids.
Chromatographic procedures currently used for the size-dependent fractionation of nucleic acids are reviewed. First, an attempt is made to clarify the concept of "size" of nucleic acids and then various aspects of the chromatography of nucleic acids are considered. It is emphasized that consideration of the dynamic three-dimensional structure of large polynucleotides in a rapidly flowing eluent is essential for both the better understanding of mechanism and the development of sophisticated procedures. Of the practical chromatographic techniques that are not based on true size fractionation, ion-exchange chromatography on non-porous column packings appears to be the most efficient. Other methods, such as hydrophobic interaction, are unlikely to become popular. As for truly size-dependent modes, there are gel permeation and slalom chromatography. Although media with extremely large pores become available, the efficiency of gel permeation is still low as a practical separation procedure for large nucleic acid molecules. Its best use seems to be in the field of physicochemical research into nucleic acids in solution. The newly discovered slalom chromatography is based on a principle completely different from all other chromatographic modes. It enables the efficient separation of large double-stranded DNA fragments of 5-50 kilo base pairs by discriminating their length. It has proved not only to be useful as a tool for nucleic acid research but also to have great significance in other fields, e.g. the hydrodynamics of polymer solutions, the search for new chromatographic modes, etc.
Kasai
K
K
Department of Biological Chemistry, Faculty of Pharmaceutical Sciences, Teikyo University, Kanagawa, Japan.
D000818:Animals; D002850:Chromatography, Gel / Q000379:methods*; D002852:Chromatography, Ion Exchange / Q000379:methods*; D006801:Humans; D009696:Nucleic Acids / Q000032:analysis* / Q000737:chemistry*
D009696:Nucleic Acids