Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Rapid Isolation of Genomic DNA from Normal and Apoptotic Cells Using Magnetic Silica Resins

  • Park, Jee-Sun (Department of Biology, College of Natural Sciences, Chungnam National University) ;
  • Park, Jung-Hyun (Protein Engineering laboratory, Korea Research Institute of Bioscience and Biotechnology) ;
  • Na, Shin-Young (Department of Biology, College of Natural Sciences, Chungnam National University) ;
  • Choe, Soo-Young (School of Life Sciences, Chungbuk National university) ;
  • Choi, Sang-Nam (Sekyung Biotechnical Research Center) ;
  • You, Kwan-Hee (Department of Biology, College of Natural Sciences, Chungnam National University)
  • Published : 2001.10.01
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Abstract

The isolation of genomic DNA from mammalian cells is usually performed by cell lysis followed by protein digestion, extraction, and finally, ethanol precipitation of the chromosomal DNA. However, in the case of large sample numbers or when only small amounts of starting materials are available, such conventional methods are not efficient and are cumbersome to be applied. Some alternative methods have been described as well as having commercial DNA isolation kits to be available, nevertheless, there is room left for much improvement. In the present study, a novel method is introduced, where it simplifies conventional protocols by omitting some time-consuming steps such as protease incubation or DNA precipitation and its resuspension. Using paramagnetic silica resins, the genomic DNA was purified over a magnetic field, and the bound DNA was eluted with a low-salt buffer. The fidelity and effectiveness of this novel method was determined by using normal and apoptotic cells as a starting material and then compared to other protocols. The high speed and convenience along with its high efficiency in detecting apoptotic chromosomal DNA will prove this method to be an improved alternative in the isolation of genomic DNA from mammalian cells.

Keywords

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