Plant Biotechnology Reports

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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Rapid and simple method for DNA extraction from plant and algal species suitable for PCR amplification using a chelating resin Chelex 100

  • HwangBo, Kwon (Plant Systems Engineering Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Son, Su-Hyun (Plant Systems Engineering Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Lee, Jong-Suk (Plant Systems Engineering Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Min, Sung-Ran (Plant Systems Engineering Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Ko, Suk-Min (Eugentech Inc., KRIBB) ;
  • Liu, Jang-R. (Plant Systems Engineering Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Choi, Dong-Su (Department of Biology, Kunsan National University) ;
  • Jeong, Won-Joong (Plant Systems Engineering Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB))
  • Received : 2009.10.29
  • Accepted : 2009.11.09
  • Published : 2010.03.30
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Abstract

A DNA extraction method using Chelex 100 is widely used for bacteria, Chlamydomonas, and animal cell lines, but only rarely for plant materials due to the need for additional time-consuming and tedious steps. We have modified the Chelex 100 protocol and successfully developed a rapid and simple method of DNA extraction for efficient PCR-based detection of transgenes from a variety of transgenic plant and algal species. Our protocol consists of homogenizing plant tissue with a pestle, boiling the homogenized tissue in a microfuge tube with 5% Chelex 100 for 5 min, and centrifuging the boiled mixture. The supernatant, which is used for PCR analysis, was able to successfully amplify transgenes in transgenic tobacco, tomato, potato, Arabidopsis, rice, strawberry, Spirodela polyrhiza, Chlamydomonas, and Porphyra tenera. The entire DNA extraction procedure requires <15 min and is therefore comparable to that used for bacteria, Chlamydomonas, and animal cell lines.

Keywords

References

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