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Frozen competent yeast cells that can be transformed with high efficiency using the LiAc/SS carrier DNA/PEG method
R Daniel Gietz1, Robert H Schiestl
1Department of Biochemistry and Medical Genetics, University of Manitoba, T250-770 Bannatyne Ave., Winnipeg, Manitoba R3E 0W3, Canada.
Nature Protocols
|April 3, 2007
Summary
This study presents an efficient protocol for creating frozen competent yeast cells. These cells can be stored and used later for high-efficiency yeast transformation, saving time for repeated experiments.
Area of Science:
- Molecular Biology
- Biotechnology
Background:
- Yeast transformation is a fundamental technique in molecular biology.
- Efficient and reliable methods for yeast transformation are crucial for research.
- Storing competent yeast cells for later use can improve experimental workflow.
Purpose of the Study:
- To develop a protocol for producing frozen competent yeast cells.
- To enable high-efficiency yeast transformation using stored cells.
- To provide a time-saving method for laboratories performing repeated yeast transformations.
Main Methods:
- The protocol involves preparing yeast cultures.
- Competent yeast cells are generated using the lithium acetate/single-stranded carrier DNA/polyethylene glycol (PEG) method.
- Cells are frozen for long-term storage.
Main Results:
- The developed protocol yields highly competent yeast cells.
- Frozen competent yeast cells can be transformed with high efficiency.
- Cell preparation and freezing take approximately 30 minutes.
Conclusions:
- This protocol offers a reliable method for producing and storing competent yeast cells.
- The use of frozen competent yeast cells streamlines the transformation process.
- This method is particularly beneficial for labs with consistent needs for specific yeast strains.
