A simple and effective separation and purification procedure for DNA fragments using Dodecyltrimethylammonium bromide

Daragh M. McLoughlin, Julie O'Brien, Jennifer J. McManus, Alexander V. Gorelov*, Kenneth A. Dawson

*Corresponding author for this work

Research output: Contribution to journalArticle (Academic Journal)peer-review

31 Citations (Scopus)

Abstract

In this work we describe a simple two step separation procedure for the separation and purification of short DNA fragments. The first step involves precipitating the DNA using the cationic surfactant dodecyltrimethylammonium bromide. Dodecyltrimethylammonium bromide, unlike cetyltrimethylammonium bromide will not precipitate DNA before complexation is complete thus providing a high purity DNA. The second step involves dissolution of the DNA-dodecyltrimethylammonium complex in 75% ethanol, followed by precipitation of the Sodium-DNA salt, by titrating in a salt solution. This method is particularly suited to purification of short fragments as it does not require high salt concentrations in the ethanol precipitation step, which can be damaging for short DNA. The ability of dodecyltrimethylammonium bromide to remove ethidium bromide from intercalation sites on the DNA is also discussed.

Original languageEnglish
Pages (from-to)307-313
Number of pages7
JournalBioseparation
Volume9
Issue number5
DOIs
Publication statusPublished - 1 Dec 2000

Keywords

  • Cationic surfactant
  • DNA
  • DTAB
  • Ethanol precipitation
  • Purification

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