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KMID : 1024220140290010007
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Environmental Health and Toxicology : EHT
2014 Volume.29 No. 1 p.7 ~ p.7
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Characterization of denaturation and renaturation of DNA for DNA hybridization
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:Xiaofang Wang
:Lim Hyun-Jeong/:Son Ah-Jeong
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Abstract
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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.
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KEYWORD
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Beads mill, Dimethyl sulfoxide, DNA denaturation, DNA renaturation, Heating, Sonication
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