We have developed a set of procedures which simultaneously solubilize, denature, and stabilize the RNA present in whole cells or in cytoplasmic extracts for direct analysis by gel electrophoresis and hybridization of blotted gels. Multiple samples of tissue culture cells can be prepared for electrophoresis in less than an hour. The number of cells that can be processed for whole cell RNA is limited only be the sensitivity of detection of specific RNAs using hybridization probes. Cell preparations with high levels of RNase, such as human lymphocytes and HL60, can be processed reliably. The method involves the stabilization of RNA in solubilized cells, cytoplasm or nuclei using varioius combinations of vandayl ribonucleoside complex (VRC), SDS, proteinase K, formaldehyde and heat, followed by resolution and specific detection of the RNA species on formaldehyde gels containing SDS.
