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Preparative isolation of polymerase chain reaction products using mixed-mode chromatography
Authors:T. Matos,G. Silva,J.A. Queiroz,L. Bü  low
Affiliation:1. Department of Pure and Applied Biochemistry, Lund University, SE-221 00 Lund, Sweden;2. Health Sciences Research Centre, University of Beira Interior, 6200 506 Covilhã, Portugal
Abstract:The polymerase chain reaction (PCR) has become one of the most useful techniques in molecular biology laboratories around the world. The purification of the target DNA product is often challenging, however, and most users are restricted to employing available commercial kits. The recent developments in mixed-mode chromatography have shown higher selectivity for a variety of nucleic acid-containing samples. Capto Adhere is a mixed-mode chromatography resin that offers a high-selectivity ligand and is here applied for the purification of amplified DNAs from PCR mixtures in a 10-min single step, with yields above 95%, high linearity, and high precision for different concentrations.
Keywords:Polymerase chain reaction   Single-stranded DNA   Double-stranded DNA   Plasmid DNA   Ion-exchange chromatography   Hydrophobic interaction chromatography
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