Separation of quantitation of intracellular forms of poliovirus RNA by agarose gel electrophoresis

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Abstract
Intracellular poliovirus-specific RNA species can be measured directly by electrophoresis of total cytoplasmic nucleic acids through 1% agarose gels, resulting in the separation of single- and double-stranded forms of poliovirus RNA from each other and from [human cervical carcinoma] HeLa cell 28S rRNA. Single-stranded RNA molecules differing by only 15% in length are resolved in this gel system. RNA species can be visualized as fluorescent bands appearing after staining of the gels with ethidium bromide and observation under UV illumination. The total amount of RNA can be determined by densitometric quantitation of the fluorescent response. Thus, the amount of poliovirus-specific RNA within the cytoplasm of HeLa cells infected for various times was estimated. At 170 min postinfection, there are 0.67 .times. 105 molecules of single-stranded poliovirus RNA/cell and at 230 min, the amount increases to 3.7 .times. 105 molecules/cell. Poliovirus double-stranded RNA reaches a maximum of 0.7 .times. 105 molecules/cell at 330 min after infection.

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