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Electrophoresis

On the kinetics of monomer incorporation into polyacrylamide gels, as investigated by capillary zone electrophoresis.


PMID 8375347

Abstract

The kinetics of monomer incorporation into a polyacrylamide gel have been studied in a photopolymerization system comprising 100 microM methylene blue in presence of a red-ox system, 1 mM sodium toluenesulfinate (reducer) and 50 microM diphenyliodonium chloride (oxidizer). A precise assessment of gel point (pc) was obtained in a droplet chamber, in which argon was gently bubbled with a fused silica capillary into the reaction mixture. At pc, 50% (+/- 3) acrylamide was incorporated into the matrix, vs. 80% (+/- 4) N,N'-methylenebisacrylamide. This incorporation level remained the same when polymerized in the 2-36 degrees C temperature range. Incorporation continued almost linearly for acrylamide up to 80% conversion. The reaction was continued up to 55 min (at 2 degrees C), at which point bisacrylamide had been essentially consumed (> 99.5% incorporation) and acrylamide had reacted (95%). At 2 degrees C, after gelation, the gel became progressively turbid (the Tyndall effect plateauing at 50 min), but it remained fully transparent if, at the gel point, reaction was continued at 50 degrees C. The consumption of the pendant double bonds of Bis followed the progression of turbidity. It is concluded that, by gelation at 2 degrees C, the nascent chains form clusters held together by hydrogen bonds (melting point at 28 degrees C); such clusters are subsequently "frozen" in the three-dimensional space as the pendant double bonds in the chains react progressively. Such turbid matrices are more porous and less elastic than when the gel is polymerized at 50 degrees C. This process is similar to the "lateral aggregation" occurring when gels are formed in presence of a polymer in solution (e.g. 10 KDa polyethylene glycol; Righetti et al., Electrophoresis 1992, 13, 587-594).

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