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

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.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

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. 1

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

  2. 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).

  3. 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).

  4. 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).

  5. 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).

  6. 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).

  7. 7

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

  8. 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).

  9. 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).

  10. 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).

  11. 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).

  12. 12

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

Download references

Author information

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

Further reading

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.