Characterization of methacrylate monoliths for purification of DNA molecules.
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.
Bencina
Mojca
M
Laboratory for Biotechnology and Industrial Mycology, National Institute of Chemistry, Hajdrihova 19, SI-1000 Ljubljana, Slovenia.
Podgornik
Ales
A
Strancar
Ales
A
D001234:Aspergillus niger / Q000378:metabolism; D002845:Chromatography; D002851:Chromatography, High Pressure Liquid; D004247:DNA / Q000032:analysis / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D006863:Hydrogen-Ion Concentration; D007477:Ions; D008689:Methacrylates / Q000737:chemistry*; D010957:Plasmids / Q000302:isolation & purification / Q000378:metabolism; D012965:Sodium Chloride / Q000494:pharmacology; D013997:Time Factors
D007477:Ions; D008689:Methacrylates; D012965:Sodium Chloride; D004247:DNA