Characterization of Denaturation and Renaturation of DNA for DNA Hybridization

Overview

Journal Environ Health Toxicol

Specialty Environmental Health

Date 2014 Sep 20

PMID 25234413

Citations 38

Authors

Xiaofang Wang

Hyun Jeong Lim

Ahjeong Son

Affiliations

Soon will be listed here.

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.

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