Open AccessCharacterization of denaturation and renaturation of DNA for DNA hybridization
Author(s) -
Xiaofang Wang,
Hyun Jeong Lim,
Ahjeong Son
Publication year - 2014
Publication title -
environmental health and toxicology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.238
H-Index - 4
ISSN - 2233-6567
DOI - 10.5620/eht.2014.29.e2014007
Subject(s) - denaturation (fissile materials) , dna , chemistry , sonication , dimethyl sulfoxide , nucleic acid denaturation , sodium hydroxide , guanidine , nucleic acid thermodynamics , biochemistry , chromatography , biophysics , organic chemistry , nuclear chemistry , biology , base sequence
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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