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Purification Of Plasmid Dna From Clarified And Non-Clarified< I> Escherichia Coli Lysates By Berenil Pseudo-Affinity Chromatography.

J Chromatogr B Analyt Technol Biomed Life Sci.. 2012-09;  904:81-7
Caramelo-Nunes C, Gabriel MF, Almeida P, Marcos JC, Tomaz CT. CICS-UBI - Health Sciences Research Centre, University of Beira Interior, Av. Infante D. Henrique, 6200-506 Covilhã, Portugal.
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摘要

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 than those previously reported for other pseudo-affinity and hydrophobic interaction chromatography based processes. Total retention of all lysate components was achieved with 1.3M ammonium sulphate in the eluent buffer and pDNA elution was obtained by decreasing the salt concentration to 0.55 M. All impur... More

关键词

Plasmid purification; Small DNA ligands; Berenil; Pseudo-affinity chromatography