UracilDNAGlycosylase(UNG)catalysesthereleaseoffreeUracilfromUracil-containingDNA.UNGefficientlyhydrolyzesuracilfromsingle-strandedordouble-strandedDNA,butnotfromoligomers(6fewerbases).UNGisactiveoverabroadpHrangewithanoptimumatpH-8.0,doesn'trequiredivalentcation,andisinhibitedbyhighionicstrength(>200mM).TheabasicsitesformedinDNAbyUNGmaybecleavedbyheat,alkali-treatmentorendonucleasesthatcleavespecificallyatabasicsites.||Source:|UracilDNAGlycosylase,fromtheE.ColistrainthatcarriestheUNGgene||Applications:|SuitableforGlycosylasemediatedsinglenucleotidepolymorphismdetection(GMPD),site-directedmutagenesis,asaprobeforprotein-DNAinteractionsudies,forrapidandefficientcloningofPCRproductsandtoeliminatecarry-overcontaminationinPCR.||10XUNGReactionBuffer(Included):|Suppliedasaliquidin200mMTris-HCl,pH8.0at25°C,10mMDTT,10mMEDTA.||UnitDefinition:|1UnitisdefinedastheamountofUDGenzymethatcatalyzesreleaseof60pmolofUracil/minutefromdouble-stranded,uracil-containingDNAinatotalreactionvolumeof50ulin30minutesat37°Cin1XUracilDNAGlycosilaseReactionBufferwith1UnitofUracilDNAGlycosylaseand0.2ug[3H]-UracilDNA(10e4-10e5cpm/ug).||StorageandStABIlity:|Maybestoredat4°Cforshort-termonly.Aliquottoavoidrepeatedfreezingandthawing.Storeat-20°C.Formaximumrecoveryofproduct,centrifugetheoriginalvialafterthawingandpriortoremovingthecap.Furtherdilutionscanbemadeinassaybuffer.

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