Bioanalytical Chemistry 8. Gel Electrophoresis and Blotting
91 Bioanalytical chemistry 8. Gel electrophoresis and blotting Suggested reading: Sections 9.1, 9.2.3, 9.2.4, 9.5.1, 10.1 to 10.7, 11.1 to 11.5, and 15.5 of Mikkelsen and Cortón, Bioanalytical Chemistry Primary Source Material • Chapter 4 and 6 of Biochemistry: Berg, Jeremy M.; Tymoczko, John L.; and Stryer, Lubert (NCBI bookshelf). • Chapter 3 and 7 of Molecular Cell Biology 4th ed. (Ch. 9, 5th ed.): Lodish, Harvey; Berk, Arnold; Zipursky, S. Lawrence; Matsudaira, Paul; Baltimore, David; Darnell, James E. (NCBI bookshelf). • Chapter 12 of Introduction to Genetic Analysis Anthony: J.F. Griffiths, Jeffrey H. Miller, David T. Suzuki, Richard C. Lewontin, William M. Gelbart (NCBI bookshelf). • Some animations are from http://www.wiley-vch.de/books/info/3-527-30300-6/. • Cancer examples from Weinberg, Robert (2007). The Biology of Cancer. Garland Science. • http://www.piercenet.com/ Electrophoresis 92 The velocity of migration (v) of a molecule in an electric field depends on the electric field strength (E), the net charge on the protein (z), and the frictional coefficient (f). Ez v = f The frictional coefficient f depends on both the mass and shape of the migrating molecule and the viscosity (η) of the medium. For a sphere of radius r, f = 6πηr € The speed of migration is therefore proportional to the charge:mass ratio. z Will the charge to mass ratio differ v ∝ between proteins? Between r different DNA molecules? € • Electrophoresis is a technique for separating, or resolving, molecules in a mixture under the influence of an applied electric field.
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