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
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Ghirlando R, Wachtel EJ, Arad T and Minsky A (1992) DNA packaging induced by micellar aggregates: A novel in vitro DNA condensation system. Biochemistry 31: 7110–7119.
Gorelov AV, Kudryashov ED, Jacquier JC, McLoughlin DM and Dawson KA (1998) Complex formation between DNA and cationic surfactant. Physica A 249: 216–225.
Gray DM, Ratliff Rl and Vaughan MR (1992) Circular dichroism spectroscopy of DNA. Methods Enzymol. 211: 389–405.
Jacquier JC, Gorelov AV, McLoughlin DM and Dawson KA (1998) Capillary electrophoretic study of the complex formation between DNA and cationic surfactants. J. Chromatogr. A 817: 263–271
Jost JP, Jiricny J and Saluz A (1989) Quantative precipitation of short oligonucleotides with low concentrations of CTAB. Nuc. Acids. Res. 17: 2143–2143.
McLoughlin DM, McManus JJ, Gorelov AV and Dawson KA (2000) DNA complexes with cationic surfactant in mixed solvents: The influence of excess surfactant and salt. Prog. Colloid. Polym. Sci. (In press).
Morimoto M, Ferchmin PA and Bennett EL (1978) Spectrophotometric analysis of RNA and DNA using cetyltrimethylammonium bromide. Anal. Biochem. 62: 436–448.
Murray MG & Thompson WF (1980) Rapid isolation of high molecular weight plant DNA. Nuc. Acids Res. 8: 4321–4325.
Pasternack RF, Caccam M, Keogh B, Stephenson TA, Williams AP and Gibbs EJ (1991) Long range fluorescence quenching of ethidium ions by cationic pophyrins in the presence of DNA. J. Am. Chem. Soc. 113: 6835–6840.
Sibantani A (1970) Precipitation of minute quantities of nucleic acids as the cetyltrimethylammonium salt. Anal. Biochem. 33: 279–285.
Trend BL, Knoll DA, Ueno DA, Evans DF and Bloomfield VA (1990) Cation radius effects on the helix to coil transition of DNA. Biophys. J. 57: 829–834.
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McLoughlin, D.M., O'Brien, J., McManus, J.J. et al. A simple and effective separation and purification procedure for DNA fragments using Dodecyltrimethylammonium bromide. Bioseparation 9, 307–313 (2000). https://doi.org/10.1023/A:1011182806783
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DOI: https://doi.org/10.1023/A:1011182806783