DNA Purification

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

2、..............168HowMuchPurityDoesYourApplicationRequire?........168HowMuchNucleicAcidCanBeProducedfromaGivenAmountofStartingMaterial?........................168DoYouRequireHighMolecularWeightMaterial?........168HowImportantIsSpeedtoYourSituation?.............168

3、HowImportantIsCost?..............................169HowImportantIsReproducibility(Robustness)oftheProcedure?.......................................169WhatInterfereswithNucleicAcidPurification?..........169WhatPracticesWillMaximizetheQualityofDNAPurification?.......

4、................................171HowCanYouMaximizetheStorageLifeofPurifiedDNA?............................................172IsolatingDNAfromCellsandTissue.....................172WhatAretheFundamentalStepsofDNAPurification?.......................................1

5、72WhatAretheStrengthsandLimitationsofContemporaryPurificationMethods?..............................174WhatAretheStepsofPlasmidPurification?.............180WhatAretheOptionsforPurificationafterInVitroReactions?............................................184SpunColumn

6、ChromatographythroughGelFiltrationResins............................................184167FilterCartridges.....................................185SilicaResin-BasedStrategies...........................186IsolationfromElectrophoresisGels....................187WhatA

7、reYourOptionsforMonitoringtheQualityofYourDNAPreparation?............................190Bibliography...........................................191WHATCRITERIACOULDYOUCONSIDERWHENSELECTINGAPURIFICATIONSTRATEGY?HowMuchPurityDoesYourApplicationRequire?Whatcontaminan

8、tswillaffectyourimmediateanddownstreamapplication(s)?AsdiscussedbelowandinChapter1,“PlanningforSuccessintheLaboratory,”timeandmoneycanbesavedbydeterminingwhich