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Acrylamide Polymerization A Practical Approach

to begin the polymerization chain reaction (Shi and Jackowski 1998). The elongating polymer chains are randomly crosslinked by bis, resulting in a gel with a characteristic porosity which depends on the polymerization conditions and monomer concentrations.

Riboflavin (or riboflavin-5'-phosphate) may also be used as a source of free radicals, often in combination with TEMED and ammonium persulfate. In the presence of light and oxygen, riboflavin is converted to its leuco form, which is active in initiating polymerization. This is usually referred to as photochemical polymerization.

Polyacrylamide Gel Polymerization


Purity of Gel-Forming Reagents
Acrylamide
Gel-forming reagents include the monomers, acrylamide and bis, as well as the initiators, usually ammonium persulfate and TEMED or, occasionally, riboflavin and TEMED. On a molar basis, acrylamide is by far the most abundant component in the monomer solution. As a result, acrylamide may be the primary source of interfering contaminants (Dirksen and Chrambach 1972). Poor-quality acrylamide contains significant amounts of the following contaminants:

1. Acrylic acid Acrylic acid is the deamidation product of acrylamide. Acrylic acid will copolymerize with acrylamide and bis, thereby conferring ion exchange properties on the resulting gel. This can lead to local pH changes in the gel and cause artifacts such as aberrant relative mobility, precipitation of some proteins and nucleic acids, streaking or smearing of bands, and run-to-run irreproducibility. In acrylamide, acrylic acid should be below 0.001% (w/w). This is determined by direct titration, and supported by both
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