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BIOL 1104 Biological Techniques, Instrumentation and Data Processing Laboratory Report Name: LAM YU CHING U. no.: 2002288759 Group: 5 Date: 11/11/02 (Monday) Experiment G: Polyacrylamide gel electrophoresis on a slab gel Objective 1. To investigate a relationship between log10 (molecular mass) and protein's Rf values. 2. To predict what protein(s) is/are mixed in protein mixture X. Introduction In this experiment (SDS-PAGE), the protein separation is only based on size, not charge, as SDS has masked the proteins with an overall negative charge. Polyacrylamide gel is prepared by polymerization of neurotoxic acylamide monomer and bis-acrylamide activated by persulfate and TEMED. By varying concentration of monomer, different pore size can be attained. To improve resolution, the sharp sample protein bands visualized by bromophenol blue, are maintained by discontinuous buffer system or disc electrophoresis, which requires a buffered 2-gel system. The 'running gel' in which separation takes place is over-layered by a short, large-pored 'stacking' gel. This system employs the principle of isotachophoresis, which effectively concentrates samples from large...

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