Transformation of Saccharomyces cerevisiae by electroporation

Abstract

A method for introducing heterologous DNA into Saccharomyces cerevisiae rapidly and efficiently by electroporeation was developed. Transformant colonies appeared somewhat sooner than by the LiCl or spheroplast transformation method, and the time spent in manipulation was much less than for these two methods. The pores in the cell membrane formed by the high voltage of electroporation were resealed within 6 to 7 min after electroporation. At a capacitance of 25 μF, the optimum voltage was 2.0 to 2.25 kV/cm. Log-phase cells concentrated to 10 to 20 units at an optical density of 600 nm in 200 μl of fresh rich medium and electroporated at 2.25 kV/cm in the presence of 0.1 μg of supercoiled plasmid DNA will yield 1,000 to 4,500 colonies per μg of DNA.