12-Electrophoresis

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1、MolecularBiologyProblemSolver:ALaboratoryGuide.EditedbyAlanS.GersteinCopyright©2001byWiley-Liss,Inc.ISBNs:0-471-37972-7(Paper);0-471-22390-5(Electronic)12ElectrophoresisMarthaL.BoozChemicalSafety.......................................334WhatIstheSafestApproachtoWorkingwithAcrylamide?..

2、....................................334WhatAretheSymptomsofAcrylamidePoisoning?......335WhatIstheMedicalResponsetoAccidentalAcrylamideExposure?........................................335HowCanYouDisposeofExcess,UnusableAcrylamide?..335WhatIstheShelfLifeofAcrylamideandAcrylamideSolution

3、s?........................................336ElectricalSafety.......................................336WhatAretheRequirementsforaSafeWorkArea?......336WhatAretheRequirementsforSafeEquipmentinGoodWorkingOrder?..................................337Polyacrylamide(PAGE)Gels—BeforeSelectinga

4、Gel:GettingtheBestResultsforYourPurpose................337WhatIstheMechanismofAcrylamidePolymerization?...338WhatOtherCrosslinkersAreAvailable,andWhenShouldTheyBeUsed?.............................338HowDoYouControlPoreSize?......................339HowDoYouCalculate%Tand%C?.............

5、......341IamgratefultoBruceGoodrichforthefigureondegassingacrylamide,toFionaLeungforthedataregardingthemolecularweightvs.relativemobilitycurve,andtoLeeOlechandDaveGarfinforfruitfuldiscussionsaboutmanyofthequestionsinthischapter.331WhyShouldYouOverlaytheGel?WhatShouldYouUseforanOverlay?..

6、.................................341RegardingReproduciblePolymerization,WhatPracticesWillEnsureThatYourBandsRuntheSameWayEveryTime?...........................................341WhatCatalystConcentrationShouldYouUse?..........343WhatIstheImportanceofReagentPurityonProteinElectrophoresis

7、andStaining?.......................343WhichGelShouldYouUse?SDS-PAGE,NativePAGEorIsoelectricFocusing?..................................345WillYourSDSGelAccuratelyIndicatetheMolecularWeightofYourProteins?...........................345ShouldYouUseaStraight%GeloraGradientGel?.....345What