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A rapid and simple procedure for high‐efficiency lithium acetate transformation of cryopreserved Schizosaccharomyces pombe cells by Minoru Suga; Toyomasa Hatakeyama is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

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## Abstract A rapid, simple, convenient, and highly efficient transformation of the fission yeast __Schizosaccharomyces pombe__ has been developed. Freezing fission yeast cells in glycerol, a permeating cryoprotectant, with lithium acetate improved remarkably the transformation efficiency by one to two orders of magnitude. The optimum concentration of glycerol was found to be 30%, which is higher than that (10–15%) in the conventional cryopreservation of yeast cells. Glycerol not only played a role in cryopreserving the competent cells but also improved the transformation efficiency of the process. The thawed cell suspension with glycerol and lithium acetate was immediately mixed with carrier DNA, plasmid DNA and polyethylene glycol. Next, the mixture was heat shocked and directly spread on a selection plate. This simple procedure yielded more than 10^6^ transformants/μg plasmid DNA, reducing the time required to only 20 min in total, including the thawing time. Furthermore, the frozen competent cells were stored long‐term for more than 3 months without any significant loss of efficiency. Copyright © 2005 John Wiley & Sons, Ltd.

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Author

Minoru Suga; Toyomasa Hatakeyama

Publisher

John Wiley and Sons; Wiley (Blackwell Publishing); John Wiley & Sons Inc.; [Chichester]; [New York]: John Wiley & Sons, ©1985-; Wiley; Hindawi Limited (ISSN 0749-503X)

Published

2005

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)