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BIOC23: Lec 7 - Gel electrophoresis.doc

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Biological Sciences
Rongmin Zhao

Lec 7 Gel Electrophoresis Gel electrophoresissimilar to ion Xchange chromatography bc bothseparate charge mocs win matrix but in electrophoresis the mocs are separated by electric fieldoseparation based on DIFFERENTIAL MIGRATION mobility win matrixo3 key factors which will affect differential migrationCharged mocsSupporting matrixElectric fieldFor a given moc the charge is often fixed but you can play around w supporting matrix and electric fieldVelocityvoltagecharge of mocfrictiondistanceoFrictional coefficient is fxn of moc radius and matrix viscosityoIf you want to increase velocity can increase voltage but may cause excess heat or change charge of moc but this is hard to manipulateoBesides that you can decrease distance between electrods or decrease viscosity of matrix to increase velocityMatricesSolid if liquid particles will diffuse and no separationSolid matricesprevent loss of resolution by diffusion and maintains the separation after electrophoresis is completePropertiesoHighly hydrophilic to prevent hydrophobic interactions btn protein and mediumoUncharged this point is diff from ion Xchange bc there the matrix is charged and stable over wised range of temp pH and osmolarityoEasy to control and adjust porosityLow voltage thin sheet electrophoresisSupporting mediumo1 Paper Made of cellulose hydrophilic absorb water and mocs canseparate on paperOnly low voltage can be used cant readily adjust porosityCapacity for resolutionlowMore diffusiono2 Cellulose AcetateSuitable for microtechniques such as separation involving immunodiffusion and immunoelectrophoresisClinical appBetter resolution than paper but still some diffusionCant easily preserve separating mocso3 Thin layer of materialthin layer electrophoresis
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