A colony procedure for transformation of Saccharomyces cerevisiae |
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Authors: | David Keszenman-Pereyra Kotaro Hieda |
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Affiliation: | (1) Biophysics Laboratory, Department of Physics, Rikkyo University, Nishi-Ikebukuro 3, 171 Toshima-ku, Tokyo, Japan |
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Abstract: | Summary A rapid and simple yeast transformation procedure has been developed using colonies on agar plates. Saccharomyces cerevisiae SHY3 cells were picked up from colonies on YPD plates grown freshly or stored at 4 °C and incubated with M13RK9-T DNA at 30 °C for 1–2 h in a solution of Li+, Ca2+, Mg2+, triacetin and polyethylene glycol. About 3,500 transformants were obtained per µg of double stranded M13RK9-T DNA. Unlike the existing spheroplast techniques, single stranded M13RK9-T DNA transformed intact cells below one-hundredth frequency of the duplex form. |
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Keywords: | Saccharomyces cerevisiae Transformation Plasmid Colony Polyethylene glycol |
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