A rapid, efficient method for isolating DNA from yeast.
A method is described for the purification of chromosomal and plasmid DNA from the yeast Saccharomyces cerevisiae. This method is rapid, gives 75% of theoretical yield, and produces DNA that can be cut with restriction endonucleases. Yeast cells are treated with zymolyase, and the resulting spheroplasts are lysed in the presence of the chaotropic agent guanidine hydrochloride. After a brief ethanol precipitation, protein is removed by treatment with proteinase K followed by phenol-chloroform extraction. After ethanol precipitation, the DNA is sufficiently pure for restriction analysis or for the transformation of Escherichia coli.
D002875:Chromosomes / Q000032:analysis; D004262:DNA Restriction Enzymes; D004271:DNA, Fungal / Q000302:isolation & purification*; D004587:Electrophoresis, Agar Gel / Q000379:methods; D007202:Indicators and Reagents; D008970:Molecular Weight; D010957:Plasmids; D011865:Radioisotope Dilution Technique; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D014316:Tritium; D014498:Uracil / Q000378:metabolism
D004271:DNA, Fungal; D007202:Indicators and Reagents; D014316:Tritium; D014498:Uracil; D004262:DNA Restriction Enzymes