Primary capture of high molecular weight nucleic acids using aqueous two-phase systems

F Luechau, TC Ling, Andrew Lyddiatt

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

The practical feasibility and generic applicability of the selective partition of plasmid DNA (pDNA) and RNA in polyethylene glycol 300 (PEG-300) and di-potassium hydrogen phosphate aqueous two-phase systems (ATPS) by addition of NaCl salt was investigated with partially disrupted Escherichia coli (E. coli) cell paste. The process yielded a relatively protein-free pDNA solution with 89.5% pDNA recovery while low molecular weight RNA was partially removed. The results gained here led the way to an alternative process options for the processing of pDNA. A reproducible process involving two extraction steps to recover pDNA from partially disrupted E. coli cell paste was developed and is presented here. (C) 2008 Elsevier B.V. All rights reserved.
Original languageEnglish
Pages (from-to)202-207
Number of pages6
JournalSeparation and Purification Technology
Volume66
Issue number1
DOIs
Publication statusPublished - 1 Apr 2009

Keywords

  • ATPS
  • Selective partition
  • Plasmid DNA
  • NaCl
  • E. coli cell paste

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