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Excellgen/T4 DNA Ligase/EG-1041/200,000 Units

DescriptionT4DNALigasecatalyzestheformationofaphosphodiesterbondbetweentheterminal5′phosphateanda3′hydroxylgroupsofduplexDNAorRNA.TheenzymeefficientlyjoinsbluntandcohesiveendsandrepairssinglestrandednicksinduplexDNA,RNAorDNA/RNAhybrids.CohesiveligationwithT4DNALigasecanbecompletedinlessthan10min.
Applications
  • Cloningofrestrictionfragments
  • Joininglinkersandadapterstoblunt-endedDNA
  • Libraryconstruction
  • TAcloning
  • RecirculizationoflinearDNA
SourcePurifiedfromarecombinantE.colistraincarryingtheT4DNALigasegene
Components
  • T4DNALigase,600,000U/mlinstoragebuffer:10mMTris-HCl,50mMKCl,1mMDTT,0.1mMEDTA,50%glycerol,pH7.5@25°C.
  • 4XT4DNALigationBuffer:250mMTris-HCI,40mMMgCl2,4mMDTT,4mMATP,30%PEG6000,pH7.6@25°C
UnitDefinition1unitisdefinedastheamountofT4DNALigaserequiredtojoin50%of100ngofDNAfragmentswithcohesiveterminiin50µl1XT4DNALigaseBufferfollowinga30minuteincubationat23°C.
QualityControl
  • Theabsenceofendo-,exodeoxyribonucleases,phosphatases,andribonucleasesconfirmedbyappropriatequalitytests.
  • Functionallytestedforthecapacitytojoincohesive-andblunt-endDNAfragments.
Storage-20°C.
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