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US 2020/0263162 A1Published application
Method for preparing electrocompetent yeast cells, and method for using said cells
AssigneeImmatics Biotechnologies GmbH
InventorsSebastian BUNK, Dominik MAURER, Felix UNVERDORBEN
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Abstract
The present invention relates to improved yeast transformation of yeast cells and yeast cell libraries transformed thereby. More specifically, the present invention relates to the transformation of yeast by electroporation.
Claims20
Electrocompetent yeast cells comprising a transformation efficiency of higher than about 1×108 transformants/μg DNA, wherein said electrocompetent yeast cells are produced by the method of
- a) growing yeast cells to an OD600 of between 1.0 to 2;
- b) washing the cells with cold water;
- c) washing the cells with a cold solution comprising sorbitol and CaCl2;
- d) incubating the cells a solution comprising lithium acetate and tris2-carboxyethyl)phosphine (TCEP);
- e) washing the cells with a cold solution comprising sorbitol and CaCl2;
- f) resuspending the cells in a solution comprising sorbitol;
- and
- g) optionally, suitably storing said cells.
5 more elements
A kit for preparing electrocompetent yeast cells, comprising
- yeast cells,
- a first solution comprising sorbitol and CaCl2 or MgCl2, and
- a second solution comprising lithium acetate (LiAc) and tris2-carboxyethyl)phosphine (TCEP).
Dependencies are inferred from the claim text.