Polymerization of Polyacrylamide Gels: Efficency and Reproducibility as a Function of Catalyst Concentrations

A method has been developed for estimation of the extent of polymerization of acrylamide by measurement of the amide-nitrogen of residual extractable acrylamide monomer. This method has been applied to the evaluation of polymerization efficiency, its reproducibility, and its dependence on the concentrations of three catalysts (persulfate, riboflavin, and TEMED) at pH 3.9 and 0°C. The effects of TEMED on gel pH, on the relative mobility of dyes and proteins, and on the “stacking limits” of multiphasic buffer systems have also been studied. These studies permit development of guidelines by which polymerization conditions can be optimized.