KOD-FX-说明书

Highefficient&HighfidelityPCRenzymeKODFX(Code.No:KFX-101)ResearchOnlyTOYOBOCO.,LTD.LifeScienceDepartmentOSAKAJAPANCodeNo.KFX-101200UX1VialStoreat-20℃Highefficient&HighfidelityPCRenzymeKODFXKODFXwasdevelopedbaseontheKODDNApolymerasefromahyperthermophilicArchaeonThermococcuskodakaraensisKOD1.KODFXshowsexcellentelongationability,efficiencyandreliabilityinadditiontohighfidelity.TheenzymesolutionofKODFXcontainstwotypesofanti-KODDNApolymeraseantibodiesinhibitthepolymeraseand3’-5’exonucleaseactivities,andthisenableshotstartPCR.FeatureofKODFX・HighfidelityThePCRerrorratioofKODFXisabout10timeslessthanthatofrTaqDNApolymerase.・ExcellentelongationabilityKODFXenablesthefollowingamplification(Maximum):40kbfromlamdaphageDNA,24kbfromhumangenomicDNA,and13.5kbfromcDNA.・HighsuccessrateThisenzymeiseffectivefortheamplificationofGC-richtargetandcrudesamples.１．Contents(200reaction)KODFX(1.0U/μl)200μl×1viral2×PCRBufferforKODFX*1.7ml×3vial2mMdNTPs1ml×2vial*2×PCRBufferforKODFXisasolutionat-20℃.２．CautionResearchuseonly.３．QualitycontrolThequalitycheckwasperformedbyanamplificationoftPAgene(24kb).07-05４．Protocol（１）Preparethereactionmixtureasfollows;For1reactionFinalconcentration2xPCRbufferforKODFX25μl1×2mMdNTPs10μl0.4mMeach10pmol/μlPrimer#11.5μl0.3μM10pmol/μlPrimer#21.5μl0.3μMTemplateDNA≧1μlGenomicDNA～200ng/50μlPlasmidDNA～50ng/50μlcDNA(from～200ngRNA)/50μlKODFX(1.0U/μl)1μl1.0U/50μlAutoclaved,distilledwaterupto50μl（２）RecommendedcycleconditionsThefollowingcyclingstepsarerecommended.Thestep-downcycleconditioniseffectiveforthetroubleshootingofasmearamplificationonalong-distancePCR.2-stepcycle3-stepcyclePredenature:94℃,2min.Predenature:94℃,2min.Denature:98℃,10sec.Denature:98℃,10sec.Extension:68℃,1min./kbAnnealing:(Tm-5)℃,30sec.Extension:68℃,1min./kbStep-downcyclePredenature:94℃，2min.Denature:98℃，10sec.Extension:74℃，1min./kbDenature:98℃，10sec.Extension:72℃，1min./kbDenature:98℃，10sec.Extension:70℃，1min./kbDenature:98℃，10sec.Extension:68℃，1min./kbExtension:68℃，7min.・Theappropriateextensiontimeis1minper1kb.25～40cycles25～40cycles5cycles5cycles5cycles15～25cyclesNOTICETOPURCHASER:LIMITEDLICENSEUseofthisproductiscoveredbyoneormoreofthefollowingUSpatentsandcorrespondingpatentclaimsoutsidetheUS:5,079,352,5,789,224,5,618,711,6,127,155andclaimsoutsidetheUScorrespondingtoUSPatentNo.4,889,818.Thepurchaseofthisproductincludesalimited,non-transferableimmunityfromsuitundertheforegoingpatentclaimsforusingonlythisamountofproductforthepurchaser’sowninternalresearch.Norightunderanyotherpatentclaim(suchasthepatented5’NucleaseProcessclaimsinUSPatentsNos.5,210,015and5,487,972),norighttoperformanypatentedmethod,andnorighttoperformcommercialservicesofanykind,includingwithoutlimitationreportingtheresultsofpurchaser'sactivitiesforafeeorothercommercialconsideration,isconveyedexpressly,byimplication,orbyestoppel.Thisproductisforresearchuseonly.DiagnosticusesunderRochepatentsrequireaseparatelicensefromRoche.FurtherinformationonpurchasinglicensesmaybeobtainedbycontactingtheDirectorofLicensing,AppliedBiosystems,850LincolnCentreDrive,FosterCity,California94404,USA.TheuseofbetaineinDNApolymerasereactionsiscoveredbyintellectualproperty,includingU.S.PatentNo.6,270,962andrelatedpatentsissuedorpendinginothercountries(I.P.),exclusivelylicensedtoEPICENTRETechnologiesCorporation,726PostRoad,Madison,WI53713,USA(EPICENTRE).TherighttousebetaineinconjunctionwithKODDNAPolymeraseinPCRissublicensedbyEPICENTREtoTOYOBOCo.,Ltd.solelyforresearchpurposes.Thepurchaseofthisproductconveystothebuyeralimited,non-exclusive,non-transferablerightundertheI.P.tousethepurchasedproductcontainingbetaineandKODDNAPolymeraseinPCRsolelyforresearchpurposes.Norightsaregrantedtoresell,repackage,orfurthersublicense.Nootherlicenseisgrantedtothebuyer,whetherexpressly,byimplication,byestoppelorotherwise.TOYOBOCO.,LTD.LifeScienceDepartment2-8,DojimaHama2-chome,Kita-ku,Osaka530-8230,JAPAN