Informace o publikaci

Chromatographic behaviour and purification of linear lambda phage and plasmid DNA molecules on 2-hydroxyethyl methacrylate-ethylene dimethacrylate-based supports



Druh Článek v odborném periodiku
Časopis / Zdroj Journal of Chromatography A
Fakulta / Pracoviště MU

Přírodovědecká fakulta

Obor Analytická chemie, separace
Klíčová slova Stationary phases; LC; slalom chromatography; DNA; hydroxyethyl methacrylate-ethylene dimethacrylate
Popis The HEMA-BIO 1000 supports based on a copolymer of 2-hydroxyethyl methacrylate and ethylene dimethacrylate were used for separation of lambda DNA and its fragments and plasmid pBR322 DNA. The separation of fragments greater than 6.6 kbp was demonstrated according to the slalom chromatography mechanism on column for gel permeation chromatography in the case of linear lambda DNA fragments. The influence of the particle size of column packing, mobile phase rate, and KCl concentration in mobile phase was discussed. The purification of plasmid DNA pBR322 using GPC was more rapid in comparison with gel electrophoresis. The presence of salts in the eluate is not disadvantageous. DNA can be recovered from the eluate by ethanol precipitation. Plasmid DNA pBR322 isolated in this way was suitable for different biological applications (cleavage with restrictases, electrotransformation into bacterial cells).
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