定量DNA斑点印迹分析：镁离子对DNA与膜结合的抑制作用。
Quantitative DNA slot blot analysis: inhibition of DNA binding to membranes by magnesium ions.
作者信息
Kube D M, Srivastava A
机构信息
Department of Microbiology and Immunology, Indiana University School of Medicine, Indianapolis, IN 46202, USA.
出版信息
Nucleic Acids Res. 1997 Aug 15;25(16):3375-6. doi: 10.1093/nar/25.16.3375.
Abstract
Titers of wild-type and recombinant adeno-associated viruses are routinely determined by DNA slot blot analysis. The binding of viral DNA to nylon membranes was found to be inhibited by magnesium ions, which are critical components of the DNase I digestion carried out prior to slot blot analysis. Mg2+ions also interfered with the adsorption of plasmid DNA to nylon and nitrocellulose membranes. These observations yield practical insights into the poorly understood mechanisms by which DNA molecules are retained on solid supports.
摘要
野生型和重组腺相关病毒的滴度通常通过DNA斑点印迹分析来测定。发现病毒DNA与尼龙膜的结合受到镁离子的抑制，而镁离子是斑点印迹分析之前进行的DNase I消化的关键成分。Mg2+离子还干扰质粒DNA对尼龙膜和硝酸纤维素膜的吸附。这些观察结果为DNA分子保留在固体支持物上这一了解甚少的机制提供了实际见解。
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