Gel electrophoretic analysis of poly(riboadenylic acid)

Abstract

It is shown that molecular weights and molecular-weight distributions of poly(rA), and by implication other single-stranded polynucleotides, and synthetic and natural polyelectrolytes in general, can be determined by electrophoresis in polyacrylamide gels. It is shown that fractions of very narrow molecular-weight distribution can be obtained by preparative electrophoresis of polydisperse samples. Molecular-weight calibrations based on sedimentation coefficients of such fractions are given, and in aqueous systems do not coincide with calibrations for partially base-paired RNA species. Poly(rU) fractions fall on the same calibration as poly(rA). Relations between mobilities, relative to standard markers, and molecular weight for poly(rA) over a wide range of molecular weights are given, which allow rapid molecular-weight determination on poly(rA) samples, such as the segments found in many types of messenger RNA.