Characterization of denaturation and renaturation of DNA for DNA hybridization
Environmental Health and Toxicology
;
: e2014007-2014.
Article
in English
| WPRIM
| ID: wpr-206476
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.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Sodium Hydroxide
/
Sonication
/
Baths
/
DNA
/
Dimethyl Sulfoxide
/
Hot Temperature
/
Heating
/
Nucleic Acid Denaturation
Language:
English
Journal:
Environmental Health and Toxicology
Year:
2014
Type:
Article
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