Environmental Analysis Health and Toxicology

The Korean Society of Environmental Health and Toxicology (환경독성보건학회)
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Characterization of denaturation and renaturation of DNA for DNA hybridization

  • Wang, Xiaofang (Department of Civil Engineering, Auburn University) ;
  • Lim, Hyun Jeong (Department of Environmental Science and Engineering, Ewha Womans University) ;
  • Son, Ahjeong (Department of Environmental Science and Engineering, Ewha Womans University)
  • Received : 2014.06.12
  • Accepted : 2014.08.08
  • Published : 2014.01.01
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Abstract

Objectives The present study was designed to systematically characterize the denaturation and the renaturation of double stranded DNA (dsDNA), which is suitable for DNA hybridization. Methods A series of physical and chemical denaturation methods were implemented on well-defined 86-bp dsDNA fragment. The degree of each denaturation was measured and the most suitable denaturation method was determined. DNA renaturation tendency was also investigated for the suggested denaturation method. Results Heating, beads mill, and sonication bath did not show any denaturation for 30 minutes. However probe sonication fully denatured DNA in 5 minutes. 1 mol/L sodium hydroxide (alkaline treatment) and 60% dimethyl sulfoxide (DMSO) treatment fully denatured DNA in 2-5 minutes. Conclusions Among all the physical methods applied, the direct probe sonication was the most effective way to denature the DNA fragments. Among chemical methods, 60% DMSO was the most adequate denaturation method since it does not cause full renaturation during DNA hybridization.

Keywords

References

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