Separation and Quantitation of Intracellular Forms of Poliovirus RNA by Agarose Gel Electrophoresis

Martinez J. Hewlett, Shmuel Rozenblatt, Victor Ambros, David Baltimore

Research output: Contribution to journalArticlepeer-review

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 HeLa cell 28S ribosomal RNA. 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 ultraviolet illumination. The total amount of RNA can be determined by densitometric quantitation of the fluorescent response. In this way, the amount of poliovirus-specific RNA within the cytoplasm of HeLa cells infected for various times has been estimated. At 170-min postinfection, there are 0.67 × 105 molecules of single-stranded poliovirus RNA per cell and at 230 min, the amount has increased to 3.7 × 105 molecules/cell. Poliovirus double-stranded RNA reaches a maximum of 0.7 × 105 molecules/cell at 330 min after infection.

Original languageEnglish
Pages (from-to)2763-2767
Number of pages5
JournalBiochemistry
Volume16
Issue number12
DOIs
StatePublished - 1 Jun 1977
Externally publishedYes

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