High-performance liquid chromatography of high-molecular-weight nucleic acids on the macroporous ion exchanger, nucleogen

Abstract The application of Nucleogen★, a series of novel silica gel-based anion exchangers of different pore size in the range 60–4000 A, to analytical and large-scale high-performance liquid chromatography of a wide variety of nucleic acids is described. The influence of pore size, pH, temperature, eluting salts, bivalent metal ions, denaturing reagents and flow-rate on resolution, purity and capacity was studied. High resolution was obtained for oligonucleotides up to a chainlength of 37, natural RNSs from tRNS up to phage RNA (molecular weight 1,000,000), double-stranded RNA and DNA fragments, and supercoiled plasmids from a crude cell lysate. Recovery was over 95%; the isolated nucleic acids were pure according to gel electrophoresis, physico-chemical criteria, and activity in enzymatic tests.

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