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

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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.

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Abbreviations

CIA:

chloroform-isoamyl alcohol

CTAB:

hexadecyltrimethyl-ammonium bromide

DIECA:

diethyldithiocarbamic acid

RAPD:

random amplified polymorphic DNA

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Author notes

  1. Andrew H. Paterson

    Present address: Department of Soil and Crop Sciences, Texas A & M University, 77843-2474, College Station, TX, USA

Authors and Affiliations

  1. Department of Soil and Crop Sciences, Texas A & M University, 77843-2474, College Station, TX, USA

    Jonathan F. Wendel

  2. Department of Botany, Iowa State University, 50011, Ames, IA, USA

    Curt L. Brubaker

Authors
  1. Andrew H. Paterson
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  2. Curt L. Brubaker
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  3. Jonathan F. Wendel
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Paterson, A.H., Brubaker, C.L. & Wendel, J.F. A rapid method for extraction of cotton (Gossypium spp.) genomic DNA suitable for RFLP or PCR analysis. Plant Mol Biol Rep 11, 122–127 (1993). https://doi.org/10.1007/BF02670470

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  • DOI: https://doi.org/10.1007/BF02670470

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