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Two-step process for initial capture of plasmid DNA and partial removal of RNA using aqueous two-phase systems

Lookup NU author(s): Professor Andy Lyddiatt

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Abstract

In this paper, a two-step process for initial capture of plasmid DNA (pDNA) and partial removal of RNA using polyethylene glycol (PEG) and di-potassium hydrogen phosphate aqueous two-phase systems (ATPS) has been investigated. A Kuhni-type ATPS extraction column was prepared with 50 ml (12% (w/w) PEG 1450, 12% (w/w) phosphate) of stationary phase and loaded with crude mobile phase (26% (w/w) PEG 1450, 4% (w/w) phosphate and 70% (w/w) lysate) at a flow rate of 6 ml min(-1) at an impeller speed of 200 rpm. The experiment was terminated after 100 min, and after complete resettling of the phases, 45 ml of stationary phase was harvested. During a subsequent second extraction step contained 18% (w/w) PEG 300 and 14% (w/w) phosphate, a proportion of RNA, which was also concentrated during the column process, was removed. It was demonstrated that the recovery of pDNA in the second bottom phase was 89.4%, which was similar to the initial recovery after column extraction (92.1%). (C) 2010 Elsevier Ltd. All rights reserved.

Publication metadata

Author(s): Luechau F, Ling TC, Lyddiatt A

Publication type: Article

Publication status: Published

Journal: Process Biochemistry

Year: 2010

Volume: 45

Issue: 8

Pages: 1432-1436

Print publication date: 11/06/2010

ISSN (print): 1359-5113

ISSN (electronic): 1873-3298

Publisher: Elsevier Ltd

URL: http://dx.doi.org/10.1016/j.procbio.2010.05.015

DOI: 10.1016/j.procbio.2010.05.015

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