Rapid and simple method for DNA extraction from plant and algal species suitable for PCR amplification using a chelating resin Chelex 100

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• 作者：Kwon HwangBo (1)
  Su Hyun Son (1) (2)
  Jong Suk Lee (1)
  Sung Ran Min (1)
  Suk Min Ko (3)
  Jang R. Liu (1)
  Dongsu Choi (2)
  Won Joong Jeong (1)
  
• 关键词：Chelex 100 ; DNA extraction method ; Plant material ; PCR ; Transgenes
• 刊名：Plant Biotechnology Reports
• 出版年：2010
• 出版时间：January 2010
• 年：2010
• 卷：4
• 期：1
• 页码：49-52
• 全文大小：216KB
• 参考文献：1. Berthold DA, Best BA, Malkin R (1993) A rapid DNA preparation for PCR from / Chlamydomonas reinhardtii and / Arabidopsis thaliana. Plant Mol Biol Rep 11:338-44 CrossRef
  2. Cao M, Fu Y, Guo Y, Pan J (2009) / Chlamydomonas (Chlorophyceae) colony PCR. Protoplasma 235:107-10 CrossRef
  3. Furukawa K, Bahavanandan VP (1983) Influences of anionic polysaccharides on DNA synthesis in isolated nuclei by DNA polymerase alpha: correlation of observed effects with properties of polysaccharides. Biochim Biophys Acta 740:466-75
  4. Holmes AR, Cannon MG, Shepherd MG (1994) Detection of / Candida albicans and other yeast in blood by PCR. J Clin Microbiol 32:228-31
  5. Panaccio M (1991) PCR based diagnosis in the presence of 8% (v/v) blood. Nucleic Acid Res 19:291-92
  6. Tebbe CC, Vahjen W (1993) Interference of humic acids and DNA extracted from soil in detection and transformation of recombinant DNA from bacteria and yeast. Appl Environ Microbiol 59:2657-665
  7. Tsai YL, Olson BH (1992) Rapid method for separation of bacterial DNA from humic substances in sediments for polymerase chain reaction. Appl Environ Microbiol 58:2292-295
  8. Walsh PS, Metzger DA, Higuchi R (1991) Chelex 100 as a medium for simple extraction of DNA for PCR-based typing from forensic material. Biotechniques 10:506-13
  9. Ward AC (1992) Rapid analysis of yeast transformants using colony-PCR. Biotechniques 13:350
  
• 作者单位：Kwon HwangBo (1)
  Su Hyun Son (1) (2)
  Jong Suk Lee (1)
  Sung Ran Min (1)
  Suk Min Ko (3)
  Jang R. Liu (1)
  Dongsu Choi (2)
  Won Joong Jeong (1)
  
  1. Plant Systems Engineering Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), 111 Gwahangno, Yuseong-gu, Daejeon, 305-806, Korea
  2. Department of Biology, Kunsan National University, Miryong-dong, Gunsan, 573-701, Korea
  3. Eugentech Inc., KRIBB, Daejeon, 305-806, Korea
  

文摘

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.