Plant Molecular Biology Reporter

, Volume 11, Issue 2, pp 122–127 | Cite as

A rapid method for extraction of cotton (Gossypium spp.) genomic DNA suitable for RFLP or PCR analysis

  • Andrew H. Paterson
  • Curt L. Brubaker
  • Jonathan F. Wendel
Commentary

Abstract

Extraction of high-quality genomic DNA fromGossypium (cotton) species is difficult due to high levels of polysaccharide, oxidizable quinones, and other interfering substances. We describe a procedure that consistently permits isolation of cotton genomic DNA of satisfactory size and quality for RFLP and PCR analysis, as well as for most routine cloning applications. Several antioxidants, phenol-binding reagents, and phenol oxidase inhibitors are used throughout the procedure, and most polysaccharides are eliminated early in the procedure by isolation of nuclei.

Key Words

cotton genomic DNA extraction Gossypium PCR RFLP 

Abbreviations

CIA

chloroform-isoamyl alcohol

CTAB

hexadecyltrimethyl-ammonium bromide

DIECA

diethyldithiocarbamic acid

RAPD

random amplified polymorphic DNA

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Bernatzky, R. B., and S. D. Tanksley. 1986. Toward a saturated linkage map in tomato based on isozymes and cDNA clones. Genetics 112:887–898.PubMedGoogle Scholar
  2. Couch, J. A., and P. Fritz. 1990. Extraction of DNA from plants high in polyphenolics. Plant Mol. Biol. Reptr. 8 (1):8–12.CrossRefGoogle Scholar
  3. Fryxell, P. A. A revised taxonomic interpretation of Gossypium. RHEEDIA, in press.Google Scholar
  4. Murray, M., and W. F. Thompson. 1980. Rapid isolation of high molecular weight plant DNA. Nucl. Acids Res. 8:4321–4325.CrossRefPubMedGoogle Scholar
  5. Katterman, F. R. H., and V. I. Shattuck. 1983. An effective method of DNA isolation from the mature leaves ofGossypium species that contain large amounts of phenolic terpenoids and tannins. Prep. Biochem. 13(4):347–359.CrossRefPubMedGoogle Scholar
  6. Loomis, W. D. 1974. Overcoming problems of phenolics and quinones in the isolation of plant enzymes and organelles. Meth. Enzymol. 31:528–545.PubMedGoogle Scholar
  7. Sambrook, J., E. F. Fritsch, and T. Maniatis. 1989.Molecular Cloning: A Laboratory Manual, 2nd ed., Cold Spring Harbor Press, Cold Spring Harbor, NY.Google Scholar
  8. Wendel, J. F. 1989. New-world tetraploid cottons contain Old World cytoplasm. Proc. Natl. Acad. Sci. USA 86:4132–4136.CrossRefPubMedGoogle Scholar
  9. Williams, J. G. K., A. R. Kubelik, K. J. Livak, J. A. Rafalski, and S. V. Tingey. 1990. Oligonucleotide primers of arbitrary sequence amplify DNA polymorphisms which are useful as genetic markers. Nucl. Acids Res. 18:6531–6535.CrossRefPubMedGoogle Scholar

Copyright information

© International Society for Plant Molecular Biology 1993

Authors and Affiliations

  • Andrew H. Paterson
    • 1
  • Curt L. Brubaker
    • 2
  • Jonathan F. Wendel
    • 1
  1. 1.Department of Soil and Crop SciencesTexas A & M UniversityCollege StationUSA
  2. 2.Department of BotanyIowa State UniversityAmesUSA

Personalised recommendations