Characterization of denaturation and renaturation of DNA for DNA hybridization.
10.5620/eht.2014.29.e2014007
- Author:
Xiaofang WANG
1
;
Hyun Jeong LIM
;
Ahjeong SON
Author Information
1. Department of Civil Engineering, Auburn University, Auburn, AL, USA.
- Publication Type:Original Article
- Keywords:
Beads mill;
Dimethyl sulfoxide;
DNA denaturation;
DNA renaturation;
Heating;
Sonication
- MeSH:
Baths;
Dimethyl Sulfoxide;
DNA*;
Heating;
Hot Temperature;
Nucleic Acid Denaturation;
Sodium Hydroxide;
Sonication
- From:Environmental Health and Toxicology
2014;29(1):e2014007-
- CountryRepublic of Korea
- Language:English
-
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.
