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[34] Preparative acrylamide gel electrophoresis: Continuous and disc techniques

1971; Academic Press; Linguagem: Inglês

10.1016/0076-6879(71)22036-x

1557-7988

Louis Shuster,

Microfluidic and Bio-sensing Technologies

This chapter discusses methods for continuous and discontinuous preparative electrophoresis. The purity of acrylamide and bisacrylamide is more important for preparative than for analytical electrophoresis because of the problems that can result from inhomogeneities or from swelling of the gel. Both monomers are neurotoxic, and contact with the dry powders or with solutions or inhalation of the dust should be avoided. These procedures should be carried out in a well-ventilated hood. Heat dissipation can be a problem in the scaling up of gel electrophoresis. The temperature rise will be greatest at the center of the gel, with the result that protein bands will be bowed downward. Many preparative columns have a cold finger in the center and an outside jacket. With a short, wide column of gel, enough cooling can be achieved by recirculating the buffers. The column may be cooled with ice water flowing at a rate of several liters a minute. In addition to a pump for circulating cooling fluid it is very useful to have one or more well-controlled peristaltic pumps. These can be used for layering water over the gel during casting, for layering the sample evenly on the surface of the gel, for recirculating the buffer among electrode chambers in a continuous system, and for elution of bands as they emerge from the gel.