Novel electrophoretic approach to study of poly(A)+RNA pool heterogeneity in different areas of human postmortem...

A simple method for fast fractionation of human postmortem brain total poly(A)+RNA has been developed. Guanidine thiocyanate extractable RNA was directly applied on the membranes of cellulose nitrate with immobilized oligo(dT) chains. After the electrophoresis in 35 mM tris-acetete buffer (pH 7.90) containing 10 mM EDTA and 35% formamide, the membranes were washed with the same buffer for removal of nonbinding poly(A)-lacking RNAs. Then, poly(A)+RNA fractions were selectively stained by 3,5-dimethylphenol in the presence of FeCl3 at pH 2.35. Finally, the resulting electrophorograms were scanned at 500 nm for quantitative evaluation of the data.