一种从酵母中分离DNA的快速、高效方法。

A rapid, efficient method for isolating DNA from yeast.

作者信息

Holm C, Meeks-Wagner D W, Fangman W L, Botstein D

出版信息

Gene. 1986;42(2):169-73. doi: 10.1016/0378-1119(86)90293-3.

Abstract

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.

摘要

描述了一种从酿酒酵母中纯化染色体DNA和质粒DNA的方法。该方法快速，产率达理论值的75%，且产生的DNA可用限制性内切酶切割。用溶菌酶处理酵母细胞，所得原生质球在离液剂盐酸胍存在下裂解。经过短暂的乙醇沉淀后，先用蛋白酶K处理，再用酚-氯仿萃取以去除蛋白质。乙醇沉淀后，所得DNA纯度足以用于限制性分析或转化大肠杆菌。