产品资料

Lucigen/CircLigase™ ssDNA Ligase/CL4111K/1,000 Units

Thermostableligasethatcatalyzestheintramolecularligation(i.e.circularization)ofssDNAtemplates

Intramolecularligationofsingle-strandedDNAendscontaining5'-phosphatesand3'-hydroxylgroups
ATP-dependent,catalyticenzyme
CircularizesssDNA>30baseslong
CompatIBLewithhightemperatureligationreactionsduetothermostABIlityofCircLigase™enzyme

Applications

ProductionofssDNAtemplatesforrolling-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.TheReactionBufferdoesnotcontain
ATPorMnCl₂whichmustbeaddedtothereactionatfinalconcentrationsof0.05mMATPand2.5mMMnCl₂.A2.5-mMsolutionofATPanda50-mMsolutionofMnCl₂areincluded.

QualityControl:CircLigasessDNALigaseisfreeofdetectablephosphatase,DNAexo-andendonuclease,andRNaseactivities.

ProductCitations

Polidoros,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/gkn1014

ORDERINFORMATION

CircLigase™ssDNALigaseisprovidedat100U/μl.

Contents:CircLigase™ssDNALigase,CircLigase™10XReactionBuffer,1mMATP,50mMMnCl₂,CircLigase™ssDNAControlOligo,SterileWater.

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Lucigen/CircLigase™ ssDNA Ligase/CL4111K/1,000 Units

Thermostableligasethatcatalyzestheintramolecularligation(i.e.circularization)ofssDNAtemplates

ORDERINFORMATION