产品说明
Thermostableligasethatcatalyzestheintramolecularligation(i.e.circularization)ofssDNAtemplatesIntramolecularligationofsingle-strandedDNAendscontaining5'-phosphatesand3'-hydroxylgroupsATP-dependent,catalyticenzymeCircularizesssDNA>30baseslongCompatIBLewithhightemperatureligationreactionsduetothermostABIlityofCircLigase™enzymeApplicationsProductionofssDNAtemplatesforrolling-circlereplicationorrolling-circletranscriptionexperiments.ProductionofssDNAtemplatesforRNApolymeraseandRNApolymeraseinhibitorassays.CircLigase™ssDNALigase*isathermostableATP-dependentligasethatcatalyzesintramolecularligation(i.e.circularization)ofssDNAtemplateshavinga5´-phosphateanda3´-hydroxylgroup.IncontrasttoT4DNALigaseandAmpligase®DNALigase,whichligateDNAendsthatareannealedadjacenttoeachotheronacomplementaryDNAsequence,CircLigasessDNALigaseligatesendsofssDNAintheabsenceofacomplementarysequence.TheenzymeisthereforeusefulformakingcircularssDNAmoleculesfromlinearssDNA.CircularssDNAmoleculescanbeusedassubstratesforrolling-circlereplicationorrolling-circletranscription.LinearssDNAof>30basesiscircularizedbyCircLigaseenzyme.Understandardreactionconditions,virtuallynolinearconcatamersorcircularconcatamersareproduced.InadditiontoitsactivityonssDNA,CircLigaseenzymealsohasactivityinligatingasingle-strandednucleicacidhavinga3´-hydroxylribonucleotideanda5´-phosphorylatedribonucleotideordeoxyribonucleotide.UnitDefinition:OneunitofCircLigaseenzymeconverts1pmolofalinear5´-phosphorylatedCircLigaseStandard55-merOligointoanexonucleaseI-resistantcircularformin1hourat60°Cunderstandardassayconditions.StorageBuffer:50%glycerolcontaining50mMTris-HCl(pH7.5),100mMNaCl,0.1mMEDTA,1mMDTT,and0.1%Triton®X-100.10XReactionBuffer:0.5MMOPS(pH7.5),0.1MKCl,50mMMgCl2,and10mMDTT.TheReactionBufferdoesnotcontainATPorMnCl2whichmustbeaddedtothereactionatfinalconcentrationsof0.05mMATPand2.5mMMnCl2.A2.5-mMsolutionofATPanda50-mMsolutionofMnCl2areincluded.QualityControl:CircLigasessDNALigaseisfreeofdetectablephosphatase,DNAexo-andendonuclease,andRNaseactivities.ProductCitationsPolidoros,A.N.etal.(2006)BioTechniques41,35.Shroff,H.etal.(2005)NanoLetters5(7),1509.Lin,C.etal.(2006)AngewandteChemie118,7699.Korlach,J.etal.(2008)Proc.Natl.Acad.Sci.USA105,1176.McArthur,M.andBibb,M.J.(2008)Proc.Natl.Acad.Sci.USA105,1020.Shroff,H.etal.(2008)BiophysicalJournal94,2179.Kuhn,H.andFrank-Kamenetskii,M.D.(2008)NucleicAcidsRes.36,e40.Murakami,T.et.al.(2008)NucleicAcidsRes.Doi:10.1093/nar/gkn1014ORDERINFORMATIONCircLigase™ssDNALigaseisprovidedat100U/μl.Contents:CircLigase™ssDNALigase,CircLigase™10XReactionBuffer,1mMATP,50mMMnCl2,CircLigase™ssDNAControlOligo,SterileWater.
美国Lucigen公司,自1998年成立至今,一直致力于生命科学领域相关科研产品的研究与开发,在分子生物学领域处于领导性地位。Lucigen公司主要开发各类用于基因克隆的试剂盒及相关产品,包括:CloneSmart®平端克隆试剂盒、BigEasy®线性克隆系统、pEZSeq™平端克隆试剂盒、ClonePlex™ AK文库构建试剂盒、DNA Terminator ®末端修复试剂盒、EconoTaq® DNA聚合酶及E.cloni感受态细胞等。Lucigen公司凭借其独到的产品技术,过硬的产品质量,良好的产品服务赢得了全球广大用户的信赖。
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