Skip to main content
SpringerLink
Log in

A New Approach to the Isolation of Genomic DNA from Yeast and Fungi: Preparation of DNA-containing Cell Envelopes and Their Use in PCR

  • Published:
Russian Journal of Bioorganic Chemistry Aims and scope Submit manuscript

Abstract

A simple and rapid procedure for the preparation of yeast and fungal DNA samples useful in PCR amplification was developed. The DNA was purified from proteins, lipids, polysaccharides, and other impurities by high-temperature extraction (in a boiling water bath) with buffer solutions containing chaotropic salts. Under these conditions, yeast and fungal cell envelopes remain unbroken and retain the original DNA and RNA that could be used for direct PCR amplification. We called the proposed PCR technique as the PCR using DNA-containing cell envelopes.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

REFERENCES

  1. Saiki, R.K., Gelfand, D.H., Stoffel, S., Scharf, S.J., Higuchi, R., Horn, G.T., Mullis, K.B., and Erlich, H.A., Science, 1988, vol. 239, pp. 487-491.

    Google Scholar 

  2. Williams, J.G.K., Kubelik, A.R., Livak, K.J., Rafalski, J.A., and Tingey, S.V., Nucleic Acids Res., 1990, vol. 18, pp. 6531-6535.

    Google Scholar 

  3. Welsh, J. and McClelland, M., Nucleic Acids Res., 1990, vol. 18, pp. 7213-7235.

    Google Scholar 

  4. Bulat, S.A., Kaboev, O.K., Mironenko, N.V., Ibatullin, F.M., Luchkina, L.A., and Suslov, A.V., Genetika (Moscow), 1992, vol. 28, pp. 20-28.

    Google Scholar 

  5. Glee, P.M., Russell, P.J., Welsch, J.A., Pratt, J.C., and Cutler, J.E., Anal. Biochem., 1987, vol. 164, pp. 207-213.

    Google Scholar 

  6. van Burik, J.-A.H., Schreckhise, R.W., White, T.C., Bowden, R.A., and Myerson, D., Medical Mycology, 1998, vol. 36, pp. 299-303.

    Google Scholar 

  7. Hoffman, C.S. and Winston, F., Gene, 1987, vol. 57, pp. 267-272.

    Google Scholar 

  8. Muller, F.M., Werner, K.E., Kasai, M., Francescon, A., Chanock, S.J., and Walsh, T.J., J. Clin. Microbiol., 1998, vol. 36, pp. 1625-1629.

    Google Scholar 

  9. Holm, C., Meeks-Wagner, D.W., Fangman, W.L., and Botstein, D., Gene, 1986, vol. 42, pp. 169-173.

    Google Scholar 

  10. Mann, W. and Jeffery, J., Anal. Biochem., 1989, vol. 178, pp. 82-87.

    Google Scholar 

  11. Philippsen, P., Stolz, A., and Scherf, C., Meth. Enzymol., 1991, vol. 194, pp. 169-182.

    Google Scholar 

  12. Polaina, J. and Adam, A.C., Nucleic Acids Res., 1991, vol. 19, p. 5443.

    Google Scholar 

  13. Miller, J.F. and Bolen, D.W., Biochem. Biophys. Res. Commun., 1978, vol. 81, pp. 610-615.

    Google Scholar 

  14. von Hippel, P.H. and Wong, K.-Y., Science, 1964, vol. 145, pp. 577-580.

    Google Scholar 

  15. Castellino, F.J. and Barker, R., Biochemistry, 1968, vol. 7, pp. 4135-4138.

    Google Scholar 

  16. Gordon, J.A., Biochemistry, 1972, vol. 11, pp. 1862-1870.

    Google Scholar 

  17. Saito, Y and Wada, A., Biopolymers, 1983, vol. 22, pp. 2123-2132.

    Google Scholar 

  18. Chirgwin, J.M., Przybyla, A.E., MacDonald, R.J., and Rutter, W.J., Biochemistry, 1979, vol. 18, pp. 5294-5299.

    Google Scholar 

  19. Boom, R., Sol, C.J., Salimans, M.M., Jansen, C.H., Wertheim-van Dillen, P.M., and van der Noordaa, J., J. Clin. Microbiol., 1990, vol. 28, pp. 495-503.

    Google Scholar 

  20. Chomczynski, P., BioTechniques, 1993, vol. 15, pp. 532-534, 536-537.

    Google Scholar 

  21. Chomczynski, P., Mackey, K., Drews, R., and Wilfinger, W., BioTechniques, 1997, vol. 22, pp. 550-553.

    Google Scholar 

  22. Bulat, S.A., Mironenko, N.V., and Zholkevich, Yu.G., Genetika (Moscow), 1995, vol. 31, pp. 315-323.

    Google Scholar 

  23. Birnboim, H.C. and Doly, J., Nucleic Acids Res., 1979, vol. 7, pp. 1531-1523.

    Google Scholar 

  24. Pichia Expression Kit, A Manual of Methods for Expression of Recombinant Proteins in Pichia Pastoris, Invitrogen Co., 1997. 146

  25. Shpakovskii, G.V. and Shematorova, E.K., Bioorg. Khim., 1998, vol. 24, pp. 933-937.

    Google Scholar 

  26. Vilgalys, R. and Hester, M., J. Bacteriol., 1990, vol. 172, pp. 4238-4246.

    Google Scholar 

  27. Liang, Q. and Richardson, T., BioTechniques, 1992, vol. 13, pp. 730-732, 735.

    Google Scholar 

  28. Chen, H.-R., Hsu, M.-T., and Cheng, S.-C., BioTechniques, 1995, vol. 19, p. 744, 746.

    Google Scholar 

  29. Haaning, J., Oxvig, C., Overgaard, M.T., and Sottrup-Jensen, L., Biochem. Mol. Biol. Int., 1997, vol. 42, pp. 169-172.

    Google Scholar 

  30. Jesnowski, R., Naering, J., and Wolf, K., Curr. Genet., 1995, vol. 27, pp. 318-319.

    Google Scholar 

  31. Ramotar, D., Mol. Cell. Biochem., 1995, vol. 145, pp. 185-187.

    Google Scholar 

  32. Wang, H., Kohalmi, S.E., and Gatler, A.J., Anal. Biochem., 1996, vol. 237, pp. 145-146.

    Google Scholar 

  33. Akada, R., Murakane, T., and Nishizava, Y., BioTechniques, 2000, vol. 28, pp. 668-670, 672.

    Google Scholar 

  34. Maniatis, T., Fritsch, E.F., and Sambrook, J., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor, NY: Cold Spring Harbor Lab., 1989, pp. 6.3-6.19.

    Google Scholar 

  35. Laemmli, U.K., Nature, 1970, vol. 227, pp. 680-685.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Russian Academy of Sciences, Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry, ul. Miklukho-Maklaya 16/10, GSP-7 Moscow, 117997, Russia

    V. N. Danilevich & E. V. Grishin

Authors
  1. V. N. Danilevich
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. E. V. Grishin
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to V. N. Danilevich.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Danilevich, V.N., Grishin, E.V. A New Approach to the Isolation of Genomic DNA from Yeast and Fungi: Preparation of DNA-containing Cell Envelopes and Their Use in PCR. Russian Journal of Bioorganic Chemistry 28, 136–146 (2002). https://doi.org/10.1023/A:1015021508321

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1015021508321

Search

Navigation