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Isolation of viral RNA using cesium chloride-ethidium bromide equilibrium sedimentation
Authors:G J Lancz  J V Scaletti  L C McLaren
Institution:Department of Microbiology, The University of New Mexico, School of Medicine, Albuquerque, New Mexico 87106 USA
Abstract:The effect of ethidium bromide (EB) on the buoyant density of reovirus RNA during equilibrium sedimentation has been investigated. The addition of the dye ethidium bromide was found to reduce the buoyant density of reovirus RNA in a Cs2SO4 gradient by a value of 0.13 to 0.15 g/cc, and provided a separation limit of 0.10 g/cc relative to the ? of marker DNA. Ethidium bromide was found also to reduce the ? of reovirus RNA to allow this RNA to band on a CsCl gradient. The separation factor between DNA and RNA on a CsCl-EB gradient was found to be 0.23 g/cc, indicating this type of gradient to be highly effective for separating the two types of polynucleotides.
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