从酵母中制备十分钟的DNA可有效释放自主质粒用于转化大肠杆菌。

A ten-minute DNA preparation from yeast efficiently releases autonomous plasmids for transformation of Escherichia coli.

作者信息

Hoffman C S, Winston F

机构信息

Department of Genetics, Harvard Medical School, Boston, MA 02115.

出版信息

Gene. 1987;57(2-3):267-72. doi: 10.1016/0378-1119(87)90131-4.

DOI:10.1016/0378-1119(87)90131-4

PMID:3319781

Abstract

A procedure for the rapid isolation of DNA from the yeast Saccharomyces cerevisiae is described. To release plasmid DNA for the transformation of Escherichia coli, cells are subjected to vortex mixing in the presence of acid-washed glass beads, Triton X-100, sodium dodecyl sulfate, phenol and chloroform. Centrifugation of this mixture separates the DNA from cellular debris. E. coli can be efficiently transformed with plasmid present in the aqueous layer without further purification of the plasmid DNA. This procedure also releases chromosomal DNA. Following two ethanol precipitations, the chromosomal DNA can be digested by restriction endonucleases and analysed by Southern blot analysis.

摘要

本文描述了一种从酿酒酵母中快速分离DNA的方法。为了释放用于转化大肠杆菌的质粒DNA，细胞在酸洗玻璃珠、吐温X-100、十二烷基硫酸钠、苯酚和氯仿存在的情况下进行涡旋混合。该混合物离心后可将DNA与细胞碎片分离。无需进一步纯化质粒DNA，水层中存在的质粒就能高效转化大肠杆菌。此方法也会释放染色体DNA。经过两次乙醇沉淀后，染色体DNA可用限制性内切酶消化，并通过Southern印迹分析进行检测。