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Chromatographic separation of lambda DNas

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RITTICH Bohuslav RACEK Tomáš ŠPANOVÁ Alena DOŠKAŘ Jiří

Rok publikování 1999
Druh Článek ve sborníku
Konference 13th Bratislava International Conference on Polymers. Separation and Characterization of Macromolecules
Fakulta / Pracoviště MU

Přírodovědecká fakulta

Citace
Obor Analytická chemie, separace
Popis The separation of DNA fragments are of increasing interest of molecular biology, gene technology and applied sciences. The most widely used methods, the polyacrylamide and agarose gel electrophoresis, have been limited to comparatively small molecules. Moreover the fragments are contamined by soluble impurities from gels, which are known to be inhibitors of DNA processing enzymes. The separation of DNA fragments larger than 20 kbp, which are important as inserts in recombinant DNA research, has been studied. An alternative method based on gel permeation chromatography (GPC) has been adopted for this purpose. In our work GPC on HEMA based sorbents (copolymer of 2-hydroxyethyl methacrylate and ethylene dimethacrylate) was used for separation of wild type lambda DNA molecules (48.5 kbp), their concatemers and mixture of lambda DNA fragments. Three peaks representing the 9.4, 23.1 and 48.5 kbp were eluted in the order of the slalom mode. The separation of lambda DNA concatemers was not achieved.
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