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Sequence-dependent DNA separation by anion-exchange high-performance liquid chromatography.

H Yamakawa·K i Higashino·O Ohara
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Abstract

High-performance liquid chromatography (HPLC) system with a new nonporous anion-exchange resin, DNA-NPR, made it possible to rapidly separate DNA fragments up to 20 kbp with high resolution. In order to further characterize this chromatographic DNA separation system, we prepared a mixture of double-stranded DNAs of constant length carrying a fully degenerated 50-bp region and analyzed their chromatographic behavior on the DNA-NPR column. The results indicated that the separation of DNA fragments on the anion-exchange HPLC was governed not only by size, but also by nucleotide sequence: even DNA fragments with the same size and the same base content could be separated on this column. Taking advantage of this characteristic feature of the anion-exchange HPLC, we could readily fractionate human cDNAs with practically acceptable recovery and high resolution. Furthermore, the combination of HPLC and gel electrophoresis realized separation of a mixture of DNA fragments in a two-dimensional pattern.

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Base SequenceCell LineChromatography, High Pressure LiquidChromatography, Ion ExchangeDNADNA Restriction EnzymesDNA, ComplementaryElectrophoresis, Gel, Two-DimensionalHumansMolecular Sequence DataJournal Article

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H Yamakawa, K i Higashino, O Ohara. 1996-09-05. Sequence-dependent DNA separation by anion-exchange high-performance liquid chromatography.. https://doi.org/10.1006/abio.1996.0354

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