QIAGEN病毒RNA提取试剂盒说明书

QIAamp®ViralRNAMiniHandbookForpurificationofviralRNAfromplasmaserumcell-freebodyfluidscell-culturesupernatantsSecondEditionDecember2005:QIAGEN®,QIAamp®,BioRobot®,MagAttract®,MinElute®,UltraSens®(QIAGENGroup);Centricon®,UltraFree®(MilliporeCorp.);Corex®(Corning,Inc.);Tween®(ICIAmericasInc.).ThePCRprocessiscoveredbyU.S.Patents4,683,195and4,683,202andforeignequivalentsownedbyHoffmann-LaRocheAG.©1999–2005QIAGEN,allrightsreserved.1030454_Qiag_HB19.12.20059:32UhrSeite2QIAampViralRNAMiniHandbook12/20053ContentsKitContents4Storage4ProductUseLimitations5ProductWarrantyandSatisfactionGuarantee5TechnicalAssistance6QualityControl6SafetyInformation6Introduction8Principle8Procedure8CellularDNAcontamination10Warningsandprecautions11Samplevolumes11Lysis11CarrierRNA12Additionofinternalcontrols12Spinandvacuumprocedures13Determinationofyield13DeterminationofviralRNAlength13EquipmentandReagentstoBeSuppliedbyUser14ImportantNotes15Preparationofreagents15HandlingofQIAampMinispincolumns18HandlingguidelinesfortheQIAvac24Plus19Centrifugation22ProtocolsPurificationofViralRNA(SpinProtocol)23PurificationofViralRNA(VacuumProtocol)26SampleConcentration30PurificationofCellular,Bacterial,orViralDNAfromUrine31TroubleshootingGuide32Appendix36OrderingInformation38QIAGENDistributors431030454_Qiag_HB19.12.20059:32UhrSeite34QIAampViralRNAMiniHandbook12/2005KitContentsQIAampViralRNAKit(50)(250)Catalogno.5290452906Numberofpreps50250QIAampMiniSpinColumns50250CollectionTubes(2ml)2001000BufferAVL*31ml5x31mlBufferAW1*(concentrate)19ml95mlBufferAW2†(concentrate)13ml66mlBufferAVE†3x2ml8x2mlCarrierRNA(polyA)310µg5x310µgHandbook11*Containschaotropicsaltwhichisanirritant.Notcompatiblewithdisinfectingreagentswhichcontainbleach.Seepage6forsafetyinformation.†Containssodiumazideasapreservative.StorageQIAampMinispincolumnsshouldbestoreddryatroomtemperature(15–25°C);storageathighertemperaturesshouldbeavoided.Allsolutionsshouldbestoredatroomtemperatureunlessotherwisestated.QIAampMinispincolumnsandallbuffersandreagentscanbestoredundertheseconditionsuntiltheexpirationdateonthekitboxwithoutshowinganyreductioninperformance.LyophilizedcarrierRNAcanbestoredatroomtemperature(15–25°C)untiltheexpirationdateonthekitbox.CarrierRNAshouldbedissolvedinBufferAVE;dissolvedcarrierRNAshouldbeimmediatelyaddedtoBufferAVLasdescribedonpage15.Thissolutionshouldbepreparedfresh,andisstableat2–8°Cforupto48hours.BufferAVL–carrierRNAdevelopsaprecipitatewhenstoredat2–8°Cthatmustberedissolvedbywarmingat80°Cbeforeuse.UnusedportionsofcarrierRNAdissolvedinBufferAVEshouldbefrozeninaliquotsat–20°C.Donotfreeze–thawthealiquotsofcarrierRNAmorethan3times.DONOTwarmBufferAVL–carrierRNAsolutionmorethan6times.DONOTincubateat80°Cformorethan5min.FrequentwarmingandextendedincubationwillcausedegradationofthecarrierRNA,leadingtoreducedrecoveryofviralRNAandeventuallyfalsenegativeRT-PCRresults,particularlywhenlow-titersamplesareused.1030454_Qiag_HB19.12.20059:32UhrSeite4QIAampViralRNAMiniHandbook12/20055ProductUseLimitationsQIAampViralRNAMiniKitsareintendedasgeneral-purposedevices.Noclaimorrepresentationisintendedfortheirusetoidentifyanyspecificorganismorforclinicaluse(diagnostic,prognostic,therapeutic,orbloodbanking).Itistheuser’sresponsibilitytovalidatetheperformanceofQIAampViralRNAMiniKitsforanyparticularuse,sincetheperformancecharacteristicsofthesekitshavenotbeenvalidatedforanyspecificorganism.QIAampViralRNAMiniKitsmaybeusedinclinicaldiagnosticlaboratorysystemsafterthelaboratoryhasvalidatedtheircompletesystemasrequiredbyCLIA'88regulationsintheU.S.orequivalentsinothercountries.Allduecareandattentionshouldbeexercisedinthehandlingoftheproducts.WerecommendallusersofQIAGENproductstoadheretotheNIHguidelinesthathavebeendevelopedforrecombinantDNAexperiments,ortootherapplicableguidelines.ProductWarrantyandSatisfactionGuaranteeQIAGENguaranteestheperformanceofallproductsinthemannerdescribedinourproductliterature.Thepurchasermustdeterminethesuitabilityoftheproductforitsparticularuse.Shouldanyproductfailtoperformsatisfactorilyduetoanyreasonotherthanmisuse,QIAGENwillreplaceitfreeofchargeorrefundthepurchaseprice.Wereservetherighttochange,alter,ormodifyanyproducttoenhanceitsperformanceanddesign.IfaQIAGEN®productdoesnotmeetyourexpectations,simplycallyourlocalTechnicalServiceDepartment.Wewillcredityouraccountorexchangetheproduct—asyouwish.SeparateconditionsapplytoQIAGENscientificinstruments,serviceproducts,andtoproductsshippedondryice.Pleaseinquireformoreinformation.AcopyofQIAGENtermsandconditionscanbeobtainedonrequest,andisalsoprovidedonthebackofourinvoices.Ifyouhavequestionsaboutproductspecificationsorperformance,pleasecallQIAGENTechnicalServicesoryourlocaldistributor(seebackcover).1030454_Qiag_HB19.12.20059:32UhrSeite56QIAampViralRNAMiniHandbook12/2005TechnicalAssistanceAtQIAGENweprideourselvesonthequalityandavailabilityofourtechnicalsupport.OurTechnicalServiceDepartmentsarestaffedbyexperiencedscientistswithextensivepracticalandtheoreticalexpertiseinmolecularbio