Pouring reproducible gradients in gels under computer control: new devices for simultaneous delivery of two independent gradients, for more flexible slope and pH range of immobilized pH gradients.

Abstract

Abstract Recently we described (Electrophoresis 5: 143-147, 1984) a new device for producing density and solute gradients, involving computer-controlled coöperation of stepmotor-driven high-precision burettes, the purpose being to substantially improve reproducibility, flexibility, and documentation of gradients used in gels and in other applications in biochemistry. Here we present the functional principle of three modified and partly alternative devices, based on simultaneous delivery of a density gradient and a non-density gradient. They provide unlimited flexibility of choice for the slope of the non-density gradient, which is stabilized by the density gradient. They seem especially useful for pouring immobilized pH gradients in gels of wide pH ranges (between pH 3.5 and 10) where localized flattening of the gradient is needed to selectively improve resolution. Used in two-dimensional electrophoretic analysis, they should considerably improve control of spot coördinates, standardization of the technique, and interlaboratory data exchange.