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Excellgen/T7 DNA Ligase/EG-1033/100,000 Units

Description

T7DNALigaseisanATP-dependentdsDNAligasefrombacteriophageT7.Itcatalyzestheformationofaphosphodiesterbondbetweena5′phosphateanda3′hydroxylterminiinduplexDNA.CohesiveendligationandnicksealingcanbeefficientlycatalyzedbyT7DNALigase.However,itislessefficientthanT4andT3DNALigasesforbluntendligationCohesiveligationwithT7DNALigasecanbecompletedinlessthan10minutes

Applications
  • CloningofDNAfragmentsgeneratedbyrestrictionenzymedigestion
  • AddinglinkersoradapterstodsDNA
  • CircularizationoflinearDNA
  • Nick-sealingindsDNA
  • Site-directedmutagenesis
  • GoldenGateassemblyofDNAfragmentssuchasTranscriptionActivator-LikeEffectorNucleases(TALEN)
SourcePurifiedfromarecombinantE.colistraincarryingtheT7DNALigasegene
Components
  • T7DNALigase,3,000,000U/mlinstoragebuffer:20mMTris-HCl,50mMNaCl,1mMDTT,0.1mMEDTA,50%glycerol,pH7.5@25°C.
  • 4XT7DNALigationBuffer:250mMTris-HCI,40mMMgCl2,4mMDTT,4mMATP,30%PEG6000,pH7.6@25°C
UnitDefinition1unitisdefinedastheamountofT7DNALigaserequiredtoligate50%of100ngDNAfragmentswithcohesiveterminiin30minutesat23°C
QualityControl
  • Theabsenceofendo-,exodeoxyribonucleases,phosphatases,andribonucleasesconfirmedbyappropriatequalitytests.
  • Functionallytestedforthecapacitytojoincohesive-andblunt-endDNAfragments.
Storage-20°C.
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