Analysis of single- and double-stranded nucleic acids on polyacrylamide and agarose gels by using glyoxal and acridine orange.

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Abstract
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.