Analysis of single-and double-stranded nucleic acids on polyacrylamide and agarose gels by using glyoxal and acridine orange.
GK McMaster, GG Carmichael - Proceedings of the …, 1977 - National Acad Sciences
GK McMaster, GG Carmichael
Proceedings of the National Academy of Sciences, 1977•National Acad SciencesWe have developed a simple and rapid system for the denaturation of nucleic acids and
their subsequent analysis by gel electrophoresis. RNA and DNA are denatured in 1 M
glyoxal (ethanedial) and 50%(vol/vol) dimethyl sulfoxide, at 50 degrees. 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 molecular weights are used as standards, accurate molecular weights for RNA are …
their subsequent analysis by gel electrophoresis. RNA and DNA are denatured in 1 M
glyoxal (ethanedial) and 50%(vol/vol) dimethyl sulfoxide, at 50 degrees. 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 molecular weights are used as standards, accurate molecular weights for RNA are …
We have developed a simple and rapid system for the denaturation of nucleic acids and their subsequent analysis by gel electrophoresis. RNA and DNA are denatured in 1 M glyoxal (ethanedial) and 50% (vol/vol) dimethyl sulfoxide, at 50 degrees. 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 molecular weights are used as standards, accurate molecular weights for RNA are obtained. Furthermore, we have employed the metachromatic stain acridine orange 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.
National Acad Sciences