Purification of cloned and genomic DNA by guanidine thiocyanate/isobutyl alcohol fractionation.
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.
Nelson
J E
JE
Department of Obstetrics and Gynecology, C. S. Mott Center for Human Growth and Development, Wayne State University School of Medicine, Detroit, Michigan 48201.
Krawetz
S A
SA
D000440:Butanols / Q000494:pharmacology; D003001:Cloning, Molecular; D004247:DNA / Q000187:drug effects / Q000235:genetics / Q000302:isolation & purification*; D004269:DNA, Bacterial / Q000187:drug effects / Q000235:genetics / Q000302:isolation & purification*; D004278:DNA, Superhelical / Q000235:genetics / Q000302:isolation & purification; D015894:Genome, Human; D006146:Guanidines / Q000494:pharmacology; D006801:Humans; D009691:Nucleic Acid Denaturation; D013861:Thiocyanates / Q000494:pharmacology
D000440:Butanols; D004269:DNA, Bacterial; D004278:DNA, Superhelical; D006146:Guanidines; D013861:Thiocyanates; C040507:isobutyl alcohol; C054436:guanidine thiocyanate; D004247:DNA