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Frozen competent yeast cells that can be transformed with high efficiency using the LiAc/SS carrier DNA/PEG method

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Abstract

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.

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Change history

  • 04 December 2008

    In the version of this article initially published, in the Reagent Setup section on p. 2, the recipe for lithium acetate (1.0 M) called for 102 g of lithium acetate dihydrate. This should be 10.2 g. The error has been corrected in the HTML and PDF versions of the article.

References

  1. Ito, H., Fukuda, Y., Murata, K. & Kimura, A. Transformation of intact yeast cells treated with alkali cations. J. Bacteriol. 153, 163–168 (1983).

    CAS  PubMed  PubMed Central  Google Scholar 

  2. Schiestl, R.H. & Gietz, R.D. High efficiency transformation of intact yeast cells using single-stranded nucleic acids as carrier. Curr. Genet. 16, 339–346 (1989).

    Article  CAS  Google Scholar 

  3. Gietz, R.D., Schiestl, R.H., Willems, A.R. & Woods, R.A. Studies on the transformation of intact yeast cells by the LiAc/SS-DNA/PEG procedure. Yeast 11, 355–360 (1995).

    Article  CAS  Google Scholar 

  4. Gietz, R.D. & Woods, R.A. Transformation of yeast by the lithium acetate/single-stranded carrier DNA/PEG method. in Methods in Microbiology: Yeast Gene Analysis Vol. 26 (eds. Brown, A.J.P. & Tuite, M.F.) 53–66 (Academic Press, San Diego, CA, 1998).

    Chapter  Google Scholar 

  5. Woods, R.A. & Gietz, R.D. Yeast transformation. in Gene Transfer Methods: Introducing DNA into Living Cells and Organisms (eds. Steel, L.F. & Norton, P.A.) 25–43 (Eaton Publishing, BioTechniques Books Division, Natick, MA, 2000).

    Google Scholar 

  6. Gietz, R.D. & Woods,, R.A. Yeast transformation. in Methods in Enzymology, Guide to Yeast Genetics and Cell Biology, Parts B and C Vol. 350 (eds. Guthrie, C. & Fink, G.R.) 87–96 (Academic Press, San Diego, CA, 2001).

    Google Scholar 

  7. Gietz, R.D. & Woods, R.A. Genetic transformation of yeast. BioTechniques 30, 816–831 (2001).

    Article  CAS  Google Scholar 

  8. Gietz, R.D., Triggs-Raine, B., Robbins, A., Graham, K.C. & Woods, R.A. Identification of proteins that interact with a protein of interest: applications of the yeast two-hybrid system. Mol. Cell. Biochem. 172, 67–79 (1997).

    Article  CAS  Google Scholar 

  9. Gietz, R.D. & Woods, R.A. Screening for protein–protein interactions in the yeast two-hybrid system. in Methods and Protocols, Methods in Molecular Biology, Vol. 185, Embryonic Stem Cells (ed. Turksen, K.) 471–486 (Humana Press, Totowa, NY, 2001).

    Google Scholar 

  10. Gietz, R.D. Yeast two-hybrid system screening. in Methods in Molecular Biology, Vol. 313, Yeast Protocols (ed. Xiao, W.) 345–371 (Humana Press, Totowa, NY, 2006).

    Google Scholar 

  11. Gietz, R.D. & Schiestl, R.H. High efficiency yeast transformation using the LiAc/SS carrier DNA/PEG method. Nat. Protocols 1, 31–34 (2007).

    Article  Google Scholar 

  12. Rose, M.D. Isolation of genes by complementation in yeast. Methods Enzymol. 152, 481–504 (1987).

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Biochemistry and Medical Genetics, University of Manitoba, T250-770 Bannatyne Ave., Winnipeg, R3E 0W3, Manitoba, Canada

    R Daniel Gietz

  2. Department of Pathology, Environmental Health and Radiation Oncology, UCLA School of Public Health and David Geffen School of Medicine, 650 Charles E. Young Drive South, Los Angeles, 90095, California, USA

    Robert H Schiestl

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  1. R Daniel Gietz
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  2. Robert H Schiestl
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Correspondence to Robert H Schiestl.

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Gietz, R., Schiestl, R. Frozen competent yeast cells that can be transformed with high efficiency using the LiAc/SS carrier DNA/PEG method. Nat Protoc 2, 1–4 (2007). https://doi.org/10.1038/nprot.2007.17

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