LALI ElectrophoresisI204_RCr

LALI ElectrophoresisI204_RCr - Basic Principle of...

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Unformatted text preview: Basic Principle of Electrophoresis  š E • 5 : & E o A U a ' b + O E ~ L ) * Electrophoresis  š E * h œ K *  U  '  +  U  '  + 1 Principal of electrophoresis Electrophoresis is the process of moving charged molecule in solution by applying an electric field across the mixture 2- - + -- - + + - + + - + +- + Electrode C ro s s s e c tio n dia g ra m o f g e l a ppa ra tus de s ig n: 3 A. Tube B. Vertical Slab C. Horizontal submarine D. Horizontal thin layer with buffer p B C An example of Electrophoresis 4 Principle of Separation According to charge According to size 5 (1) According to charge When charged molecules are placed in an electric field, they migrate toward either the positive (anode) or negative (cathode) pole according to their charge. 6 Mice run faster through the forest than elephant 7 1 2 (2) According to size Instruments and Reagents Buffer Power supply Supporting media Detection and Quantification 8 I. Buffer Function of buffer 1. carries the applied current 2. established the pH 3. determine the electric charge on the solute High ionic strength of buffer produce sharper band produce more heat Commonly used buffer Barbital buffer & Tris-EDTA for protein Tris-acetate-EDTA & Tris-borate-EDTA (50mmol/L; pH 7.5- 7.8) 9 II. Power Supply Function: To supply electric power Constant voltage (increasing of heat over time) Heat = E x I x t Constant current according to Ohm s law E= I x R R decrease over time, which inturn decrease the heat effect 10 E= EMF in volts (V) I = Current in amperes (A) t = Time in second (s) III.Support materials used in electrophoresis Paper Starch Agar/agarose Cellulose acetate polyacrylamide gel 11 Support Media (ii)...
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This note was uploaded on 01/07/2011 for the course CHEM 6153 taught by Professor - during the Spring '10 term at University of Florida.

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LALI ElectrophoresisI204_RCr - Basic Principle of...

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