Abstract
Purification of messenger RNA and mRNA precursors with columns of oligo (dT)-cellulose or poly(U)-Sepharose results in a highly enriched but nevertheless impure population of polyadenylated molecules. Since the contaminating molecules, principally rRNA, are present in variable but often significant amounts, methods for quantifying the polyadenylated component are required. This chapter presents a technique for determining the mole fraction poly(A)/sup +/ RNA and its molar concentration in the presence of roughly equivalent amounts of poly(A)/sup -/ RNA. The method consumes about 75-100 ng of total RNA and takes about 45 min to perform.
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