Gietz R Daniel, Woods Robin A
Department of Biochemistry and Medical Genetics, University of Manitoba, Winnipeg, Manitoba, Canada R3M 0T5.
Methods Enzymol. 2002;350:87-96. doi: 10.1016/s0076-6879(02)50957-5.
In this chapter we have provided instructions for transforming yeast by a number of variations of the LiAc/SS-DNA/PEG method for a number of different applications. The rapid transformation protocol is used when small numbers of transformants are required. The high efficiency transformation protocol is used to generate large numbers of transformants or to deliver DNA constructs or oligonucleotides into the yeast cell. The large-scale transformation protocol is primarily applicable to the analysis of complex plasmid DNA libraries, such as those required for the yeast two-hybrid system. The microtiter plate versions of the rapid and high efficiency transformation protocols can be applied to high-throughput screening technologies.
在本章中,我们针对多种不同应用,提供了通过LiAc/SS-DNA/PEG方法的多种变体来转化酵母的说明。当需要少量转化子时,使用快速转化方案。高效转化方案用于产生大量转化子,或将DNA构建体或寡核苷酸导入酵母细胞。大规模转化方案主要适用于复杂质粒DNA文库的分析,例如酵母双杂交系统所需的文库。快速和高效转化方案的微量滴定板版本可应用于高通量筛选技术。