Hofmeister series salts enhance purification of plasmid DNA by non-ionic detergents

George Lezin, Michael R. Kuehn, Luca Brunelli

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Ion-exchange chromatography is the standard technique used for plasmid DNA purification, an essential molecular biology procedure. Non-ionic detergents (NIDs) have been used for plasmid DNA purification, but it is unclear whether Hofmeister series salts (HSS) change the solubility and phase separation properties of specific NIDs, enhancing plasmid DNA purification. After scaling-up NID-mediated plasmid DNA isolation, we established that NIDs in HSS solutions minimize plasmid DNA contamination with protein. In addition, large-scale NID/HSS solutions eliminated lipopolysaccharides (LPS) contamination of plasmid DNA more effectively than Qiagen ion-exchange columns. Large-scale NID isolation/NID purification generated increased yields of high-quality DNA compared to alkali isolation/column purification. This work characterizes how HSS enhance NID-mediated plasmid DNA purification, and demonstrates that NID phase transition is not necessary for LPS removal from plasmid DNA. Specific NIDs such as IGEPAL CA-520 can be utilized for rapid, inexpensive, and efficient laboratory-based large-scale plasmid DNA purification, outperforming Qiagen-based column procedures.

Original languageEnglish (US)
Pages (from-to)1872-1882
Number of pages11
JournalBiotechnology and Bioengineering
Volume108
Issue number8
DOIs
StatePublished - Aug 1 2011

Fingerprint

Detergents
Purification
Plasmids
DNA
Salts
Distillation columns
DNA Contamination
Lipopolysaccharides
Ion exchange
Contamination
Molecular biology
Ion Exchange
Phase Transition
Ion Exchange Chromatography
Alkalies
Chromatography
Phase separation
Solubility
Molecular Biology
Phase transitions

Keywords

  • Hofmeister salts
  • Non-ionic detergents
  • Plasmid DNA purification

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

Cite this

Hofmeister series salts enhance purification of plasmid DNA by non-ionic detergents. / Lezin, George; Kuehn, Michael R.; Brunelli, Luca.

In: Biotechnology and Bioengineering, Vol. 108, No. 8, 01.08.2011, p. 1872-1882.

Research output: Contribution to journalArticle

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