TOPO Cloning and Gateway Technology.pdf

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1、TOPOTACloning®KitswithGateway®TechnologyAccelerateyourresearchwithTOPO®CloningandGateway®TechnologyWiththisforward-movingcombinationyou’ll:•Achieve5-minuteTOPOTACloning®reactionsyielding95%recombinants•Eliminatetedious,time-consumingcloning/sub-cloningsteps•Bereadyforallyour

2、downstreamresearchneedsFastentrytoaworldofdiscoveryTOPO®Cloningisthefastest,mosteffectivecloningtechnologyavailable.Gateway®Technologyprovidesastraightforward,rapidmethodforflexibleproteinexpressionandfunctionalanalysisstudies.Combiningthesetwotechnologiesquicklysatisfiesyou

3、rresearchneedstodayandsetsyouupforfuturedown-streamanalysis—evenifyou’renotsurewhereyourresearchwilltakeyoutomorrow.UseTOPO®Cloningforfast,efficientPCRcloningandGateway®TechnologyforrapidaccesstoanyofyourdownstreamapplicationsTOPOIGenomicsapplicationsProteomicsapplicationsRI

4、CCCTTAGGGREcoGGGATTCCCEcoTOPOTOPO®Gateway®attL1CloningSequencingrecombinationGeneexpression1attTL22TArchivingDNAsamplesProteinfunctionTOPO®TAiroCGateway®vectorULocalizationpStructuralanalysisinmycSpectinoUniquevectordesignkeepsyoumovingFastTOPO®CloningandflexibleGateway®Tech

5、nologyareFigure1—ThepCR®8/GW/TOPO®vectorcombinedinthepCR®8/GW/TOPO®vector(Figure1).ItisTOPOsuppliedlinearizedandactivatedwithtopoisomeraseIforeasy,ICCCTTAGGGR5-minuteligations.Inaddition,pCR®8/GW/TOPO®offers:EcoRIGGGATTCCCEco•Novelprimersiteslocatedlessthan55basepairsfromTOP

6、OthePCRproductinsertionsiteforconvenientsequencingattL11att•SpectinomycinresistancegeneforrobustselectioninE.coliTL22•EcoRIsitesflankingthePCRproductinsertionsiteforTeasyexcisionofinserts•attLsitesforrapidrecombinationintoanyGateway®®®destinationvectorforyourdownstreamapplic

7、ationsiropCR8/GW/TOPOCU2.8kbYou’llgetwhatyouneednowforyourgenomicsstudiesandbepreadyforthenextstepsyourresearchrequires.inmycSpectino28009556288TOPOTACloning®KitswithGateway®TechnologyThreesimpleTOPO®CloningstepsgetyouinTheTOPO®Cloningmethodreplacesligasewithtopoi-thentransf

8、ormE.coli(Figure2).WithTOPO®CloningsomeraseItoclonePCRproductsandotherDNAmo