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Wheat Rust Diseases pp 207–213Cite as

Isolation of Wheat Genomic DNA for Gene Mapping and Cloning

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1659))

Abstract

DNA is widely used in plant genetic and molecular biology studies. In this chapter, we describe how to extract DNA from wheat tissues. The tissue samples are ground to disrupt the cell wall. Then cetyltrimethylammonium bromide (CTAB) or sodium dodecyl sulfate (SDS) is used to disrupt the cell and nuclear membranes to release the DNA into solution. A reducing agent, β-mercaptoethanol, is added to break the disulfide bonds between the cysteine residues and to help remove the tanins and polyphenols. A high concentration of salt is employed to remove polysaccharides. Ethylenediaminetetraacetic acid (EDTA) stops DNase activity by chelating the magnesium ions. The nucleic acid solution is extracted with chloroform–isoamyl alcohol (24:1) or 6 M ammonium acetate. The DNA in aqueous phase is precipated with ethanol or isopropanol, which makes DNA less hydrophilic in the presence of sodium ions (Na+).

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References

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Author information

Authors and Affiliations

  1. John Innes Centre, Norwich Research Park, Norwich, NR4 7UH, UK

    Guotai Yu, Asyraf Hatta & Brande B. H. Wulff

  2. Department of Agriculture Technology, Universiti Putra Malaysia, Serdang, Malaysia

    Asyraf Hatta

  3. CSIRO Agriculture and Food, Canberra, ACT, Australia

    Evans Lagudah

  4. Research School of Biology, Australian National University, Canberra, ACT, Australia

    Sambasivam Periyannan

  5. Queensland Alliance for Agriculture and Food Innovation, University of Queensland, Brisbane, QLD, Australia

    Sambasivam Periyannan

  6. Plant Breeding Institute, University of Sydney, Sydney, NSW, Australia

    Sambasivam Periyannan

  7. Agriculture & Food, CSIRO, Canberra, ACT, Australia

    Sambasivam Periyannan

Authors
  1. Guotai Yu
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  2. Asyraf Hatta
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  3. Sambasivam Periyannan
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  4. Evans Lagudah
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  5. Brande B. H. Wulff
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Corresponding author

Correspondence to Brande B. H. Wulff .

Editor information

Editors and Affiliations

  1. Research School of Biology, Australian National University, Canberra, Aust Capital Terr, Australia

    Sambasivam Periyannan

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© 2017 Springer Science+Business Media LLC

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Yu, G., Hatta, A., Periyannan, S., Lagudah, E., Wulff, B.B.H. (2017). Isolation of Wheat Genomic DNA for Gene Mapping and Cloning. In: Periyannan, S. (eds) Wheat Rust Diseases. Methods in Molecular Biology, vol 1659. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7249-4_18

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  • DOI: https://doi.org/10.1007/978-1-4939-7249-4_18

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7248-7

  • Online ISBN: 978-1-4939-7249-4

  • eBook Packages: Springer Protocols

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