Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Protocol
  • Published:

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

This article has been updated

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.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Prices may be subject to local taxes which are calculated during checkout

Similar content being viewed by others

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 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Robert H Schiestl.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Rights and permissions

Reprints and permissions

About this article

Cite this article

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

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nprot.2007.17

This article is cited by

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing