Gietz R Daniel, Schiestl Robert H
Department of Biochemistry and Medical Genetics, University of Manitoba, T250-770 Bannatyne Ave., Winnipeg, Manitoba R3E 0W3, Canada.
Nat Protoc. 2007;2(1):1-4. doi: 10.1038/nprot.2007.17.
Here we describe a protocol for the production of frozen competent yeast cells that can be transformed with high efficiency using the lithium acetate/single-stranded carrier DNA/PEG method. This protocol allows the production of highly competent yeast cells that can be frozen and used at a later date and is especially useful for laboratories using one or two strains repeatedly. The production of yeast cells for freezing takes only approximately 30 min, once the yeast culture has grown up. Transformation with frozen competent yeast cells will take approximately 30 min, depending on the heat shock used.
在此，我们描述了一种制备冷冻感受态酵母细胞的方法，该方法可使用醋酸锂/单链载体DNA/聚乙二醇法进行高效转化。此方法能够制备出可冷冻并在日后使用的高感受态酵母细胞，对于反复使用一两种菌株的实验室尤为有用。一旦酵母培养物生长起来，制备用于冷冻的酵母细胞仅需约30分钟。使用冷冻感受态酵母细胞进行转化，根据所采用的热激条件不同，大约需要30分钟。
