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Purification of plasmid DNA from clarified and non-clarified Escherichia coli l...
Nunes, C. Caramelo; Gabriel, M. F.; Almeida, Paulo; Marcos, João Carlos; Tomaz, C. T.In this study, berenil was tested as a ligand, specifically to purify plasmids of different sizes pVAX1-LacZ (6.05 Kbp) and pCAMBIA-1303 (12.361 Kbp) from clarified Escherichia coli alkaline lysates. For this purpose, chromatographic experiments were performed using Sepharose derivatized with berenil. The results showed that both pDNA molecules are completely purified using lower amounts of salt in the eluent t...
Specific berenil–DNA interactions : an approach for separation of plasmid isofo...
Caramelo-Nunes, C.; Tente, T.; Almeida, P.; Marcos, João Carlos; Tomaz, C. T.Small molecules, like some antibiotics and anticancer agents that bind DNA with high specificity can represent a relevant alternative as ligands in affinity processes for plasmid DNA (pDNA) purification. In the present study, pDNA binding affinities of berberine, berenil, kanamycin and neomycin were evaluated by a competitive displacement assay with ethidium bromide using a fluorimetric titration technique. The...
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