Use of critical point polyacrylamide sols in thermal denaturation experiments with chromatin at physiological ionic strength

Abstract

Low percentage highly crosslinked polyacrylamide gels just above the critical point in the chemically polymerized sol to gel transition are used to generate polyacrylamide sols at critical point concentrations, 7.4 g liter −1, by mild heating. We find that chromatin samples mixed with these sols induce the sol to gel transition in a process of complex coacervation. In this state, salt insoluble chicken erythrocyte chromatin is stabilized against large scale aggregation and precipitation during thermal denaturation at physiological sodium ion concentrations. The hyperchromic melting behavior of DNA in polyacrylamide sols is reproducible and consistent throughout a wide range of sodium chloride concentrations. Empirical spectroscopic techniques are discussed which isolate temperature-dependent hyperchromic signals at 260 nm due to conformation changes of DNA in chromatin and local environmental changes which promote anomalous light scattering.