Purification of cloned and genomic DNA by guanidine thiocyanate/isobutyl alcohol fractionation

Analytical Biochemistry
J E Nelson, S A Krawetz


We have developed a rapid and efficient method to purify cloned or high molecular weight eukaryotic DNA from cell lysates using guanidine thiocyanate and isobutyl alcohol. This simple two-step extraction procedure utilizes isobutyl alcohol to fractionate DNA away from cell lysates containing guanidine thiocyanate, yielding a relatively large quantity of pure DNA. The yield of DNA is approximately twice that obtained by the organic extraction method. DNA purified with this chaotropic-based protocol is suitable for a variety of applications including restriction analysis, double-stranded sequencing, PCR, subcloning, and transcription.


Nov 24, 1979·Nucleic Acids Research·H C Birnboim, J Doly
Jun 1, 1987·American Journal of Medical Genetics·L MadisenM E Hodes
Jan 1, 1986·Journal of Biochemical and Biophysical Methods·S A KrawetzG H Dixon
Dec 28, 1983·Biochemical and Biophysical Research Communications·S J GargerL K Grill
Jul 1, 1982·Proceedings of the National Academy of Sciences of the United States of America·D DiMaioT Maniatis


Oct 12, 2012·Mammalian Genome : Official Journal of the International Mammalian Genome Society·Carolina P GóesAndrea C Peripato
Sep 23, 1997·Journal of Virological Methods·A M ChachulskaF Vilaine
Oct 18, 2001·International Journal of Food Microbiology·A A Alsallami, R Kotłowski
Mar 1, 1997·Genetic Analysis : Biomolecular Engineering·I B ChungS A Krawetz
Jun 25, 1993·Nucleic Acids Research·C A Koller, V Kohli
Mar 14, 2014·Mitochondrial DNA. Part A. DNA Mapping, Sequencing, and Analysis·Yuttamol MuangkramSaowaphang Sanannu
Jul 30, 2014·Forensic Science International. Genetics·Waradee BuddhakosaiWorawidh Wajjwalku
Mar 28, 2002·DNA Sequence : the Journal of DNA Sequencing and Mapping·R P MartinsS A Krawetz
Dec 1, 2005·BioTechniques·Lixin ZhangCarl F Marrs
Apr 23, 2009·Molecular Oncology·Nancy BretschneiderStefanie Denger
Mar 7, 2008·Nature·Raphaël MétivierGilles Salbert
Mar 7, 2008·Nature·Sara KangaspeskaGeorge Reid
Jun 13, 1997·The Journal of Biological Chemistry·N HaradaM Morikawa
Sep 13, 2017·Development Genes and Evolution·Janaina Lima de OliveiraReinaldo Alves de Brito

Related Concepts

Isobutyl alcohol, titanium (+4) salt
Guanidine thiocyanate
DNA, Double-Stranded
DNA, Bacterial
DNA, Superhelical
RNA Denaturation
Genome, Human

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