Novel Electrophoretic Approach to Study of Poly(A)⁺RNA Pool Heterogeneity in Different Areas of Human Postmortem Brain

Abstract

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 Hacking 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.

Key Words:

• human brain poly(A)⁺ RNA
• electrophoresis
• oligo(dT)-cellulose nitrate membranes