Polyacrylamide Gel Electrophoresis
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چکیده
Polyacrylamide gels are typically formed by polymerization of the monomer acrylamide crosslinked to the co-monomer, N,N’-methylenebis-acrylamide, commonly called BIS. This process is a freeradical polymerization that requires an initiator, usually ammonium persulfate, and a catalyst like N,N,N’,N’-tetraethylmethylenediamine (TEMED). A distinct advantage of polyacrylamide gel systems is that by changing the initial concentrations of acrylamide and BIS, one can control the amount of crosslinking and hardness of the gel. The amount of crosslinking of a gel determines the size of the pores in the matrix, which in turn, controls the size of the molecules that can pass through the gel. So as the concentration (%) of acrylamide in the gel increases, the pore sizes in the gel get smaller and larger molecules are excluded from migrating through the gel. Typical acrylamide concentrations range from 3% to 20%. While agarose gels have large pore sizes ideal for separating DNA, RNA and large proteins, polyacrylamide gels have smaller pores that are better for separating small proteins, nucleic acids, polypeptides, and DNA fragments, ranging in size from 5 to 300 kDa .
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