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Plant Molecular Biology Reporter

, Volume 17, Issue 1, pp 53–57 | Cite as

A Rapid DNA Minipreparation Method Suitable for AFLP and Other PCR Applications

  • D.-H. Chen
  • P.C. Ronald
Article

Abstract

A rapid DNA minipreparation method was developed for rice and other plant species. This method uses an Eppendorf tube and 1-ml pipette tip to grind plant tissues, and requires only one transfer for DNA isolation. In a single day, one person can complete DNA isolation from more than 120 leaf samples. The yields of the DNA samples ranged from 2.3 to 5.2 μg from 25–50 mg fresh leaf tissue. DNA samples extracted using this method from rice were completely digested with five restriction enzymes (EcoR I, EcoR V, Hind III, Mse I and Pst I) and were successfully used for AFLP and other PCR applications.

CTAB DNA extraction 

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References

  1. Büschges R, Hollricher K, Panstruga R, Simons G, Wolter M, Frijters A, van Daelen R, van der Lee T, Diergaarde P, Groenendijk J, Topsch S, Vos P, Salamini F and Schuize-Lefert P (1997) The barley Mlo gene: A novel control element of plant pathogen resistance. Cell 88: 695–705.Google Scholar
  2. Chen D, Wang GL and Ronald PC (1997) Location of the rice blast resistance locus Pi5(t) in Moroberekan by AFLP bulk segregant analysis. Rice Genetics Newsletter 14: 95–98.Google Scholar
  3. Doyle JJ and Doyle JL (1990) Isolation of plant DNA from fresh tissue. Focus 12: 13–15.Google Scholar
  4. Stewart CN Jr and Via LE (1993) A rapid CTAB DNA isolation technique useful for RAPD fingerprinting and other PCR applications. BioTechniques 14: 748–758.Google Scholar
  5. Song W-Y, Wang G-L, Chen L, Kim H-S, Pi L-Y, Gardner J, Wang B, Holsten T, Zhai W-X, Zhu L-H, Fauquet C and Ronald PC (1995) A receptor kinase-like protein encoded by the rice disease resistance gene Xa21. Science 270: 1804–1806.Google Scholar
  6. Vos P, Hogers R, Bleeker M, Rijans M, van der Lee T, Hornes M, Frijters A, Pot J, Peleman J, Kuiper M and Zabeau M (1995) A new technique for DNA fingerprinting. Nucl Acid Res 23: 4407–4414.Google Scholar
  7. Wang G-L, Ruan D-L, Song W-Y, Sideris S, Chen L, Pi L-Y, Zhang S, Zhang Z, Fauquet C, Gaut BS, Hhalen MC and Ronald PC (1998) Xa21D encodes a receptor-like molecule with a leucine-rich repeat domain that determines race-specific recognition and is subject to adaptive evolution. Plant Cell 10: 765–779.Google Scholar
  8. Williams C and Ronald PC (1994) Rapid, homogenization-free isolation of rice DNA for PCR. Nucl Acids Res 22: 1917–1918.Google Scholar
  9. Zheng K, Subudhi PK, Domingo J, Magpantay G and Huang N (1995). Rapid DNA isolation for marker assisted selection in rice breeding. Rice Genetics Newsletter 12: 255–258.Google Scholar

Copyright information

© Kluwer Academic Publishers 1999

Authors and Affiliations

  • D.-H. Chen
    • 1
  • P.C. Ronald
    • 1
  1. 1.Department of Plant PathologyUniversity of CaliforniaDavis USA

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