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AgilentTechnologiesAgilentRNA6000PicoKitGuideNotices©AgilentTechnologies,Inc.2001,2003-2008,2013Nopartofthismanualmaybereproducedinanyformorbyanymeans(includingelec-tronicstorageandretrievalortranslationintoaforeignlanguage)withoutprioragree-mentandwrittenconsentfromAgilentTechnologies,Inc.asgovernedbyUnitedStatesandinternationalcopyrightlaws.ManualPartNumberG2938-90046Rev.BEdition11/2013PrintedinGermanyAgilentTechnologiesHewlett-Packard-Strasse876337WaldbronnResarchUseOnlyNotforuseinDiagnosticProcedures.WarrantyThematerialcontainedinthisdocu-mentisprovided“asis,”andissub-jecttobeingchanged,withoutnotice,infutureeditions.Further,tothemax-imumextentpermittedbyapplicablelaw,Agilentdisclaimsallwarranties,eitherexpressorimplied,withregardtothismanualandanyinformationcontainedherein,includingbutnotlimitedtotheimpliedwarrantiesofmerchantabilityandfitnessforapar-ticularpurpose.Agilentshallnotbeliableforerrorsorforincidentalorconsequentialdamagesinconnectionwiththefurnishing,use,orperfor-manceofthisdocumentorofanyinformationcontainedherein.ShouldAgilentandtheuserhaveaseparatewrittenagreementwithwarrantytermscoveringthematerialinthisdocumentthatconflictwiththeseterms,thewarrantytermsinthesep-arateagreementshallcontrol.TechnologyLicensesThehardwareand/orsoftwaredescribedinthisdocumentarefurnishedunderalicenseandmaybeusedorcopiedonlyinaccor-dancewiththetermsofsuchlicense.RestrictedRightsLegendIfsoftwareisforuseintheperformanceofaU.S.Governmentprimecontractorsubcon-tract,Softwareisdeliveredandlicensedas“Commercialcomputersoftware”asdefinedinDFAR252.227-7014(June1995),orasa“commercialitem”asdefinedinFAR2.101(a)oras“Restrictedcomputersoft-ware”asdefinedinFAR52.227-19(June1987)oranyequivalentagencyregulationorcontractclause.Use,duplicationordis-closureofSoftwareissubjecttoAgilentTechnologies’standardcommerciallicenseterms,andnon-DODDepartmentsandAgenciesoftheU.S.GovernmentwillreceivenogreaterthanRestrictedRightsasdefinedinFAR52.227-19(c)(1-2)(June1987).U.S.GovernmentuserswillreceivenogreaterthanLimitedRightsasdefinedinFAR52.227-14(June1987)orDFAR252.227-7015(b)(2)(November1995),asapplicableinanytechnicaldata.SafetyNoticesCAUTIONACAUTIONnoticedenotesahazard.Itcallsattentiontoanoperatingprocedure,practice,orthelikethat,ifnotcorrectlyper-formedoradheredto,couldresultindamagetotheproductorlossofimportantdata.DonotproceedbeyondaCAUTIONnoticeuntiltheindicatedcondi-tionsarefullyunderstoodandmet.WARNINGAWARNINGnoticedenotesahazard.Itcallsattentiontoanoperatingprocedure,practice,orthelikethat,ifnotcorrectlyperformedoradheredto,couldresultinpersonalinjuryordeath.DonotproceedbeyondaWARNINGnoticeuntiltheindi-catedconditionsarefullyunder-stoodandmet.Caliper®,LabChip®andtheLab-ChiplogoareregisteredtrademarksofCaliperTechnologiesCorp.intheU.S.andothercountries.AdobeandAcrobatareU.S.registeredtrademarksofAdobeSystemsIncorporated.Microsoft®isaU.S.registeredtrademarkofMicrosoftCorporation.SoftwareRevisionThisguideisvalidfor2100expertB.02.04andhigherrevisionoftheAgilentexpertsoftware,where04referstominorrevisionsofthesoftwarethatdonotaffectthetechnicalaccuracyofthisguide.3ContentsContents1AgilentRNA6000PicoKit42RequiredEquipmentforRNA6000PicoAssay6EquipmentSuppliedwiththeAgilent2100Bioanalyzer6AdditionalMaterialRequired(NotSupplied)63SettinguptheAssayEquipmentandBioanalyzer7SettinguptheAssayEquipmentandBioanalyzer7SettinguptheChipPrimingStation8SettinguptheBioanalyzer9VortexMixer9Startingthe2100ExpertSoftware104EssentialMeasurementPractices115PreparingtheRNALadderafterArrival136AgilentRNA6000PicoAssayProtocol14AgilentRNA6000PicoAssayProtocol14CleaningtheElectrodesbeforeRunningAssays14PreparingtheGel15PreparingtheGel-DyeMix16LoadingtheGel-DyeMix17LoadingtheRNA6000PicoConditioningSolutionandMarker18LoadingtheDilutedLadderandSamples19InsertingaChipintheAgilent2100Bioanalyzer20StartingtheChipRun21CleaningElectrodesafteraRNA6000PicoChipRun23DecontaminatingtheElectrodes247CheckingYourAgilentRNA6000PicoAssayResults25RNA6000PicoLadderWellResults25RNA6000PicoSampleWellResults26KitGuide4AgilentTechnologiesAgilentRNA6000PicoKit1Table1AgilentRNA6000PicoKit(reordernumber5067-1513)AgilentRNA6000PicoChips25RNAPicoChips3ElectrodeCleanersAgilentRNA6000PicoReagents(reordernumber5067-1514)(blue)RNA6000PicoDyeConcentrate11“ThisproductisprovidedunderalicensebyLifeTechnologiesCorporationtoAgilentTechnolo-gies.Thepurchaseofthisproductconveystothebuyerthenon-transferablerighttousethepur-chasedamountoftheproductandcomponentsoftheproductonlyasdescribedinaccompanyingproductliterature.Thesaleofthisproductisexpresslyconditionedonthebuyernotusingtheproductoritscomponents(1)inmanufacturing;(2)toprovideaservice,information,ordatatoanunaffiliatedthirdpartyforpayment;(3)fortherapeutic,diagnosticorprophylacticpurposes;(4)toresell,sellorotherwisetransferthisproductoritscomponentstoanythirdparty,oruseforanyuseotherthanuseinthesubfieldsofresearchanddevelopment,qualitycontrol,forensics,envi-ronmentalanalysis,biodefenseorfoodsafetytesting.Forinformationonpurchasingalicensetothisp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