Description: Uracil-DNAGlycosylasecatalyzesthehydrolysisoftheN-glycosylicbondbetweentheuracilandsugar,leavinganabasicsiteinuracil-containingsingleordouble-strandedDNA.Theenzymeshowsnomeasurableactivityonshortoligonucleotides(<6bases),orRNAsubstrates.
Application: -Controlofcarry-overcontaminationinPCR(1) -Glycosylasemediatedsinglenucleotidepolymorphismdetection(GMPD)(2) -Site-directedmutagenesis(3) -Asaprobeforprotein-DNAinteractionstudies(4) -SNPgenotyping -CloningofPCRproducts(5) -GenerationofsinglestrandoverhangsofPCRproductsandCDNA
Source: ArecombinantE.colistraincarryingtheUracilDNAGlycosylasegenefromE.coliK-12.
SpecificActivity:77,000U/mg
SuppliedWith: 10XUDGReactionBuffer Incubateat37°C
10XUDGReactionBuffer: 200mMTris-HCl 10mMDTT 10mMEDTA pH8.0@25°C
SuppliedIn: 10mMTris-HCl 50mMNaCl 1mMDTT 0.1mMEDTA 50%glycerol pH7.5@25°C
UnitDefinition: 1unitisdefinedastheamountofenzymethatcatalyzesthereleaseof1.8nmolofUracilin30minutesfromdouble-stranded,tritiated,Uracilcontaining-DNAat37°Cin1XUDGReactionBuffer.
RecommendedStorageCondition:-20°C
References: 1.M.C.Longoetal.,UseofuracilDNAglycosylasetocontrolcarry-overcontaminationinpolymerasechainreactions.Gene.93,125-128(1990). 2.P.Vanghan,T.V.McCarthy,AnovelprocessformutationdetectionusinguracilDNAglycosylase.NucleicAcidsRes.26,810-815(1998). 3.T.A.Kunkel,Rapidandefficientsite-specificmutagenesiswithoutphenotypicselection.Proc.Natl.Acad.Sci.USA.82,488-492(1985). 4.P.R.Devchand,etal.,Uracil-DNAglycosylaseasaprobeforprotein-DNAinteractions.NucleicAcidsRes.21,3437-3443(1993). 5.P.M.Boothetal.,AssemblyandcloningofcodingsequencesforneurotrophicfactorsdirectlyfromgenomicDNAusingpolymerasechainreactionanduracilDNAglycosylase.Gene.146,303-308(1994). |