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Copyright2014PerkinElmerHealthSciences,Inc.Allrightsreserved.PNCLS140158,Rev.ADNAHighSensitivityAssayUserGuideForLabChipGX/GXIITouch1PNCLS140158,Rev.ADNAHighSensitivityAssayUserGuideContentsSpecifications.........................................................................................................2AssaySpecifications..........................................................................................2SampleConditions.............................................................................................3KitContents.......................................................................................................4SafetyWarningsandPrecautions..........................................................................5PreparationProcedures.........................................................................................6AdditionalItemsRequired..................................................................................6PreparingtheGel-DyeSolution..........................................................................6PreparingtheDNASamples,DNALadderandtheBufferTube..........................7PreparingtheChip.............................................................................................9InsertingaChipintotheLabChipGX/GXIITouchInstrument...........................11RunningtheAssay...........................................................................................13StoringtheChip...............................................................................................16ChipCartridgeCleaning...................................................................................17Results..................................................................................................................18HTDNAHighSensitivityLadderResult............................................................18Troubleshooting...................................................................................................19FrequentlyAskedQuestions................................................................................25LabChipKitEssentialPractices...........................................................................26General............................................................................................................26Reagents..........................................................................................................27Chips................................................................................................................27Samples...........................................................................................................30ChipWellAspirationUsingaVacuum.................................................................31CustomerTechnicalSupport...............................................................................32LicensesandRightsofUse.................................................................................33Specifications2PNCLS140158,Rev.ADNAHighSensitivityAssayUserGuideSpecificationsAssaySpecifications11.AllspecificationspertainingtoDNAfragmentsweredeterminedusingladderassampleinTEbuffer.AllspecificationspertainingtoDNAsmearsweredeterminedusingCovarisshearedcontrolgenomicDNA(humanmale)inTEbuffer.Shearingtimewas30sor240s.Table1.AssaySpecificationsSizingRange50-5000bpSizingResolutiona±5%from100-500bp±10%from50-100bp,500-1000bp±15%from1000-3000bp±22%from3000-5000bpSizingAccuracy±10%SizingPrecision5%CVLinearConcentrationRangeStandardSampleWorkflow10pg/µL-500pg/µLperfragmentfrom50bpto2000bp50pg/µL-500pg/µLperfragmentfrom2000bpto5000bp100pg/µL-5ng/µLforsmearsLimitedSampleWorkflow(initialconcentration)20pg/µL-500pg/µLperfragmentfrom50bpto2000bp100pg/µL-500pg/µLperfragmentfrom2000bpto5000bp200pg/µL-5ng/µLforsmearsStandardWorkflow5pg/µLperfragment100pg/µLforsmearsLimitedSampleWorkflow(initialconcentration)10pg/µLperfragment200pg/µLforsmearsMaximumTotalDNAConcentration5ng/µLtotal,500pg/µLperfragmentQuantitationAccuracy±30%Specifications3PNCLS140158,Rev.ADNAHighSensitivityAssayUserGuideSampleConditionsQuantitationPrecision20%CVCarryover0.25%AnalysisTime68secondspersample(~2.5hoursfor96samples)NumberofSamplesperChipPrep96samplesChipLifetimeb400samples(four96-wellplatesorone384-wellplate)a.Resolutionisdefinedashalfheightorbetterseparationoftwopeaks.Actualseparationperformancecandependonthesampleandapplication.Peaksthatareresolvedlessthanhalfheightcanstillbeaccuratelyidentifiedbythesystemsoftware.b.ExpectedchiplifetimeisbasedonuseundernormallaboratoryconditionsandadherencetoPerkinElmerHealthSciences,Inc.preparationprotocols,sampleguidelinesandstorageconditions.Individualresultsmayvary.Table1.AssaySpecifications(Continued)SizingRange50-5000bpTable2.SampleConditionsAdditivesPerkinElmerHealthSciences,Inc.recommendsthatBSAanddetergentsexceeding0.05mg/mLand0.01%(v/v)respectivelyinconcentrationnotbeused.Higherconcentrationscanresultinchipfailure.Inaddition,non-aqueoussolventsarenotcompatiblewithDNALabChipproto
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