Benzonase®NucleaseDescriptionBenzonaseNuclease,Purity>99%10KU70664-3BenzonaseNucleaseHC,Purity>99%25KU71206-3BenzonaseNuclease,Purity>90%10KU70746-32.5KU70746-4BenzonaseNucleaseHC,Purity>90%25KU71205-3BenzonaseisageneticallyengineeredendonucleasefromSerratiamarcescens(1–2).TheenzymeisproducedandpurifiedfromE.colistrainW3110,amutantofstrainK12,containingtheproprietarypNUC1productionplasmid(3–4).Structurally,theproteinisadimerofidentical245aminoacid,~30kDasubunitswithtwoessentialdisulfidebonds(5–8).ThispromiscuousendonucleaseattacksanddegradesallformsofDNAandRNA(singlestranded,doublestranded,linearandcircular)andiseffectiveoverawiderangeofoperatingconditions(9).Theenzymecompletelydigestsnucleicacidsto5'-monophosphateterminatedoligonucleotides2–5basesinlength(2,10).Althoughthenucleaseiscapableofcleavageatnearlyallpositionsalonganucleicacidchain,sequence-dependentpreferenceshavebeendemonstrated(11).TheenzymeprefersGC-richregionsindsDNAwhileavoidingd(A)/d(T)-tracts.BenzonaseisavailablefromNovagenintwogrades,Purity>99%andPurity>90%.Bothpreparationspossessexceptionallyhighspecificactivitiesandaresuppliedfreefrommeasurableproteaseactivitiesandviralcontaminants.Bothgradesareavailableat25U/µloratahighconcentration(HC)of250U/µl.Benzonaseisidealforawidevarietyofapplicationswherecompletedigestionofnucleicacidsisdesirable.UnitdefinitionOneunitofBenzonaseNucleaseisdefinedastheamountofenzymethatcausesa∆A260of1.0in30min,whichcorrespondstocompletedigestionof37µgofDNA.Notethat1KU=1000units.StorageBenzonaseNucleaseissuppliedin50%glycerolcontaining50mMTris-HClpH8.0,20mMNaCland2mMMgCl2.Theenzymepreparationisstablefor2yearswhenstoredat–20°C.DONOTstoreat–70°CbecausefreezingBenzonaseNucleaseresultsinlossofactivity.©2003Novagen®,abrandofEMDBiosciences,Inc.,anaffiliateofMerckKGaA,Darmstadt,Germany.Allrightsreserved.BugBuster,FRETWorks,Lysonase,PopCulture,rLysozyme,S•Tag,theNovagenlogoandnamearetrademarksofNovagen,Inc.Benzonaseisatrad