用于DNA杂交的DNA变性与复性的表征
Characterization of denaturation and renaturation of DNA for DNA hybridization.
作者信息
Wang Xiaofang, Lim Hyun Jeong, Son Ahjeong
机构信息
Department of Civil Engineering, Auburn University, Auburn, AL, USA.
Department of Environmental Science and Engineering, Ewha Womans University, Seoul, Korea.
出版信息
Environ Health Toxicol. 2014 Sep 11;29:e2014007. doi: 10.5620/eht.2014.29.e2014007. eCollection 2014.
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.
目的
本研究旨在系统地表征适用于DNA杂交的双链DNA（dsDNA）的变性和复性。
方法
对明确界定的86碱基对双链DNA片段实施一系列物理和化学变性方法。测量每种变性程度并确定最合适的变性方法。还针对建议的变性方法研究了DNA复性趋势。
结果
加热、珠磨和超声浴在30分钟内未显示任何变性。然而，探针超声处理在5分钟内使DNA完全变性。1摩尔/升氢氧化钠（碱处理）和60%二甲基亚砜（DMSO）处理在2 - 5分钟内使DNA完全变性。
结论
在所有应用的物理方法中，直接探针超声处理是使DNA片段变性的最有效方法。在化学方法中，60%DMSO是最适当的变性方法，因为它在DNA杂交过程中不会导致完全复性。
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