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High-efficiency yeast transformation 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
PubMed
Summary

This study presents an optimized lithium acetate/single-stranded carrier DNA/polyethylene glycol (PEG) method for Saccharomyces cerevisiae transformation, achieving high efficiency and yield. This protocol is suitable for most yeast transformation needs.

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Area of Science:

  • Molecular Biology
  • Yeast Genetics

Background:

  • Transformation is crucial for genetic manipulation of Saccharomyces cerevisiae.
  • Existing methods vary in efficiency and speed.

Purpose of the Study:

  • To describe a high-efficiency transformation protocol for Saccharomyces cerevisiae.
  • To optimize the lithium acetate/single-stranded carrier DNA/PEG method.

Main Methods:

  • Utilized the lithium acetate/single-stranded carrier DNA/polyethylene glycol (PEG) transformation technique.
  • Optimized incubation times and heat shock duration.

Main Results:

  • Achieved the highest reported efficiency and yield of transformants.
  • Protocol duration is approximately 1.5 hours post-culture growth.

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Conclusions:

  • The optimized method offers superior efficiency and yield for Saccharomyces cerevisiae transformation.
  • This protocol is versatile and applicable to most transformation requirements.