Modifications of RNase protection assay for neuroscience applications

This report describes how certain modifications of the Ribonuclease (RNase) protection assay may increase its efficiency by decreasing the time and co st of the procedure without compromising reliability. We show that, under t he experimental conditions tested, the RNA samples can be precipitated by a solution of Tri Reagent in ethanol immediately following the RNase digesti on step. Drying the samples under vacuum before dissolving them in the gel loading buffer improves the consistency of the assay as compared to air dry ing. Although these modifications are applicable to the RNase protection as say in general, we present an example that used a multiprobe set we develop ed and have used effectively in the analysis of cytokine mRNA regulation in the brain. (C) 1998 Elsevier Science B.V. All rights reserved.