The influence of RNA concentration, pH, temperature, buffer, and gel concentration on the fractionation and mobility of RNA in agar gel electrophoresis was studied. The effect of these factors is consistent with the idea that the main mechanisms by which RNA fractionation is affected are change in the viscosity of the liquid phase of the gel, deviation of the molecules from the straight path due to hits with the gel strands, and desaggrepation of RNA complexes held together by very weak forces. A linear relationship was found between mobility and square root of gel concentration. © 1969.