首页 > Lifesci > Lifesci/RNase H in Glycerol/RNHG-1000/1000 u

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Lifesci/RNase H in Glycerol/RNHG-1000/1000 u

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RNHG-1000

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ProductDescriptionRibonucleaseHfromE.coliRibonucleaseHisanendonucleasewhichspecificallydegradestheRNAmoietyofaDNA:RNAhybrid,withoutaffectingtheunhybridizedRNA.Itdoesnotaffectdouble-strandedRNAorDNA.Itisprovidedina50%glycerolstoragebuffer.ItisalsoavailableinaTrehaloseBuffer(Suitableforlyophilization).Concentration1000U/mL.Pleaseinquireforcustomconcentrationsandbulkquantities.Applications:Enablingthesynthesisofsecond-strandCDNAbyremovaloftheRNAUsedinconjunctionwithAMVRTandT7RNAPolymeraseinamplificationofRNAUnitDefinition:OneunitofRNaseHisthatamountofenzymewhichcatalysistheproductionofonenmolofacid-solublenucleotidein20minutesat37ºCusingthefollowingreactionconditions:40mMTrisHCl,pH7.51.0µM[3H]-poly(rA):24µMpoly(dT)4.0mMMgCl21.0mMDTT30µg/mLBSA4.0%glycerolStorageBufferConditionsofRNaseHinGlycerol:20:0mMTrisHCl,pH7.5300mMKCl20.0mMMagnesiumAcetate7.0mMEDTA1.0mMDithiothreitol50%Glycerol0.2%TritonX100StorageBufferConditionsofRNaseHinTrehalose.Sameasabove,exceptfor1.0MTrehaloseinsteadofGlycerol.QualityControl:DNaseActivity:One-halfµgofHaefragmentsofPhiX-174DNAisincubatedat37ºCwith2.5unitsofRNaseHfor3hours,andthenelectrophoresedinanativeagarosegelsimultaneouslywithcontrolpositiveDNase1reactions.Nomorethantheequivalentof2.5X10E-4unitofDNase1isdetected.RibonucleaseActivity:OnemicrogramofanRNALadderisincubatedfor2hoursat37ºCwith4.0unitsofRNaseH,andthenelectrophoresedinanativeagarosegelsimultaneouslywithcontrolpositiveRNase1Areactions.Nomorethantheequivalentof8X10-8unitofRNase1Aisdetected.SpecificActivity:ThespecificactivityoftheE.coliRNaseHisnolessthan300,000unitspermg.References:Sambrook,J.,Fritsch,E.F.,andManiatis,T.(1989)MolecularCloning:ALaboratoryManual,(2ndEd.),8.64–8.65