4.5 Article

Characterization of methacrylate monoliths for purification of DNA molecules

Journal

JOURNAL OF SEPARATION SCIENCE
Volume 27, Issue 10-11, Pages 801-810

Publisher

WILEY-V C H VERLAG GMBH
DOI: 10.1002/jssc.200401784

Keywords

DNA purification; monoliths; chromatography; plasmid DNA; genomic DNA

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The suitability of methacrylate based anion exchange monolithic supports for the separation and purification of plasmid and genomic DNA has been explored. The effect of the size of the channels, ionic strength of the solution, and ligand density on the dynamic binding capacity has been investigated. The dynamic binding capacity was found to be flow independent, at least up to a linear velocity of 700 cm h(-1), and exceeded 9 mg mL(-1) for all types of DNA. The recovery depends on the pH value of the mobile phase and its ionic strength as well as on the density of the active groups. Under optimal conditions recoveries exceeding 80% were obtained even for genomic DNA. Finally, the suitability of this approach is demonstrated by purification of a real-life sample.

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