Skip to main content
SpringerLink
Log in

Modification of a CTAB DNA extraction protocol for plants containing high polysaccharide and polyphenol components

  • Commentary
  • Published:
Plant Molecular Biology Reporter Aims and scope Submit manuscript

Abstract

A relatively quick, inexpensive and consistent protocol for extraction of DNA from expanded leaf material containing large quantities of polyphenols, tannins and polysaccharides is described. Mature strawberry leaves, which contain high levels of these secondary components, were used as a study group. The method involves a modified CTAB extraction, employing high salt concentrations to remove polysaccharides, the use of polyvinyl pyrrolidone (PVP) to remove polyphenols, an extended RNase treatment and a phenol-chloroform extraction. Average yields range from 20 to 84 μg/g mature leaf tissue for both wild and cultivated octoploid and diploidFragaria species. Results from 60 plants were examined, and were consistently amplifiable in the RAPD reaction with as little as 0.5 ng DNA per 25-μL reaction. Presently, this is the first procedure for the isolation of DNA from mature strawberry leaf tissue that produces consistent results for a variety of different species, both octoploid and diploid, and is both stable and PCR amplifiable before and after extended storage. This procedure may prove useful for other difficult species in the family Rosaceae.

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

Abbreviations

CTAB:

hexadecyltrimethylammonium bromide

EtOH:

ethanol

PVP:

polyvinylpyrrolidone

References

  • Collins, G.G. and R.H. Symons. 1992. Extraction of nuclear DNA from grape vine leaves by a modified procedure. Plant Mol. Biol. Rept. 10:233–235.

    CAS  Google Scholar 

  • Davis, T.M., H. Yu and K.M. Haigis and P.J. McGowan. 1995. Template mixing: A method of enhancing detection and interpretation of codominant RAPD markers. Theor. Appl. Genet. 91:582–588.

    Article  CAS  Google Scholar 

  • Dellaporta, S.L., J. Wood and J.B. Hicks. 1985. Maize DNA miniprep, p. 36–37, inMolecular Biology of Plants, Malberg, J. Messing and I. Sussex, eds Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.

    Google Scholar 

  • Doyle J.J. and J.L. Doyle. 1987. A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem. Bull 19:11–15.

    Google Scholar 

  • Fang, G., S. Hammar and R. Grumet. 1992. A quick and inexpensive method for removing polysaccharides from plant genomic DNA. Biofeedback 13 (1):52–54.

    CAS  Google Scholar 

  • LaRoche, J. 1992. An easy and efficient procedure for isolating plant DNA using the Elu-Quik DNA purification kit. Sequences 36:3–4.

    Google Scholar 

  • Lodhi, M.A., G.-N. Ye, N.F. Weeden, B.I. Reisch. 1994. A simple and efficient method for DNA extractiosn from grapevine cultivars and Vitis species. Plant Mol. Biol. Rept. 12:6–13.

    CAS  Google Scholar 

  • Maliyakal, E.J. 1992. An efficient method for isolation of RNA and DNA from plants containing polyphenolics. Nucleic Acid Research 20:2381.

    Article  Google Scholar 

  • Mauro, M.-C., M. Strefeler, N.F. Weeden and B.I. Reisch. 1992. Genetic analysis of restriction fragment length polymorphisms inVitis. J. Heredity 83:18–21.

    CAS  Google Scholar 

  • Oard, J.H. and S. Dronavalli. 1992. Rapid isolation of rice and maize DNA for analysis by random-primer PCR. Plant Mol. Biol. Rept. 10:236–241.

    CAS  Google Scholar 

  • Richards E., M. Reichardt and S. Rogers. 1994. Preparation of genomic DNA from plant tissue.Current Protocols in Molecular Biology. Wiley Interscience.

  • Saghai-Maroof, M.A., K.M. Soliman, R.A. Jorgensen and R.W. Allard. 1984. Ribosomal DNA spacer-length polymorphism in barley: Mendelian inheritance, chromosomal location, and population dynamics. Proc. Natl. Acad. Sci. USA. 81:8014–8019.

    Article  PubMed  CAS  Google Scholar 

  • Sigma Chemical Company. 1993.Biochemical Organic Compounds for Research and Diagnostic Reagents. Sigma Diagnostics. St. Louis, USA. 2256 pp.

    Google Scholar 

  • Varadarajan, G.S. and C.S. Prakash. 1991. A rapid and efficient method for the extraction of total DNA from the sweet potato and its related species. Plant Mol. Biol. Rept. 9(1):6–12.

    CAS  Google Scholar 

  • Wang, H.M. Qi and A. Cutler. 1993. A simple method of preparing plant samples for PCR. Nucl. Acid Res. 21:4153–4154.

    Article  CAS  Google Scholar 

  • Webb, D.M. and S.J. Knapp. 1990. DNA extraction from a previously recalcitrant plant genus. Plant Mol. Biol. Rept. 8:180–185.

    Article  CAS  Google Scholar 

  • Williams, J.G.K., A.R. Kubelik, K.J. Livak, J.A. Rafalski and S.V. Tingey. 1990. DNA polymorphism amplified by arbitrary primers are useful genetic markers. Nucl. Acids Res. 18:6531–6535.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Agriculture Canada, Eastern Cereal and Oilseed Research Centre, K1A 0C6, Ottawa, Ontario, Canada

    Sue Porebski, L. Grant Bailey & Bernard R. Baum

Authors
  1. Sue Porebski
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. L. Grant Bailey
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Bernard R. Baum
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Porebski, S., Bailey, L.G. & Baum, B.R. Modification of a CTAB DNA extraction protocol for plants containing high polysaccharide and polyphenol components. Plant Mol Biol Rep 15, 8–15 (1997). https://doi.org/10.1007/BF02772108

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02772108

Key Words

Search

Navigation