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Improved Manufacture and Application of an Agarose Magnetizable Solid-Phase Support

Davies, M.J., Smethurst, D.E., Howard, K.M., Todd, M., Higgins, L.M., Bruce, Ian J. (1997) Improved Manufacture and Application of an Agarose Magnetizable Solid-Phase Support. Applied Biochemistry and Biotechnology - Part A Enzyme Engineering and Biotechnology, 68 (1-2). pp. 95-112. ISSN 0273-2289. (doi:10.1007/BF02785983) (The full text of this publication is not currently available from this repository. You may be able to access a copy if URLs are provided) (KAR id:71982)

The full text of this publication is not currently available from this repository. You may be able to access a copy if URLs are provided. (Contact us about this Publication)
Official URL
http://dx.doi.org/10.1007/BF02785983

Abstract

A simple, semiautomated, nonhazardous procedure for the production of a magnetizable solid-phase support (MSPS) has been developed based on the extrusion of molten agarose-iron oxide mixtures, which enables manufacture of a range of differently sized spherical agarose-iron oxide beads. This system has enabled scale-up of an original manufacture procedure and reproducible preparation of kg quantities of MSPS suitable for biomolecular purifications. An improved protocol for the isolation of plasmid DNA directly from cell lysates using this MSPS, derivatized with diethylaminoethyl (DEAE) groups, is reported. This involves a modified alkaline lysis, followed by adsorption to and elution from the support, yielding plasmid DNA of a purity comparable with, or better than, other methods of plasmid isolation. Using the same procedure, plasmid DNA can be isolated from bacterial cell culture volumes of 1.5 mL and 100 mL with equal efficiency and purity.

Item Type: Article
DOI/Identification number: 10.1007/BF02785983
Uncontrolled keywords: Bacteria (microorganisms), bacterial DNA; ferric ion; ferric oxide; microsphere; sepharose, agar gel electrophoresis; article; Escherichia coli; gel chromatography; genetics; isolation and purification; magnetism; methodology; plasmid, Chromatography, Gel; DNA, Bacterial; Electrophoresis, Agar Gel; Escherichia coli; Ferric Compounds; Magnetics; Microspheres; Plasmids; Sepharose
Divisions: Faculties > Sciences > School of Physical Sciences
Faculties > Sciences > School of Physical Sciences > Functional Materials Group
Depositing User: I. Bruce
Date Deposited: 31 Jan 2019 07:32 UTC
Last Modified: 30 May 2019 08:49 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/71982 (The current URI for this page, for reference purposes)
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