ANALYSIS OF SINGLE-STRANDED AND DOUBLE-STRANDED NUCLEIC-ACIDS ON POLYACRYLAMIDE AND AGAROSE GELS BY USING GLYOXAL AND ACRIDINE-ORANGE
ANALYSIS OF SINGLE-STRANDED AND DOUBLE-STRANDED NUCLEIC-ACIDS ON POLYACRYLAMIDE AND AGAROSE GELS BY USING GLYOXAL AND ACRIDINE-ORANGE
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DOI：
10.1073/pnas.74.11.4835
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发表时间：
1977-01-01
影响因子：
11.1
通讯作者：
CARMICHAEL, GG
中科院分区：
文献类型：
--
作者：
MCMASTER, GK;CARMICHAEL, GG
开发了一种简单快速的核酸变性及其随后的凝胶电泳分析系统。RNA和DNA在1 M乙二醛（乙醛）和50%（体积/体积）二甲基亚砜中于50 ℃变性。然后，在pH 7.0的10 mM磷酸钠缓冲液中，通过丙烯酰胺或琼脂糖凝胶对糖苷酸化的核酸进行电泳。当使用已知MW的甘氨酸化DNA分子作为标准品时，可获得RNA的准确MW。异染染料吖啶橙子用于凝胶中核酸的可视化。这种染料与双链和单链多核苷酸的相互作用不同，分别发出绿色和红色荧光。通过使用这些技术，可以在同一平板凝胶上分析天然和变性的DNA和RNA分子。
A simple and rapid system for the denaturation of nucleic acids and their subsequent analysis by gel electrophoresis was developed. RNA and DNA are denatured in 1 M glyoxal(ethanedial) and 50% (vol/vol) dimethyl sulfoxide, at 50.degree.. The glyoxalated nucleic acids are then subjected to electrophoresis through either acrylamide or agarose gels in a 10 mM sodium phosphate buffer at pH 7.0. When glyoxalated DNA molecules of known MW are used as standards, accurate MW for RNA are obtained. The metachromatic stain acridine orange was employed for visualization of nucleic acids in gels. This dye interacts differently with double- and single-stranded polynucleotides, fluorescing green and red, respectively. By using these techniques, native and denatured DNA and RNA molecules can be analyzed on the same slab gel.