Description: T3DNALigasejoinsbluntendsandcohesiveendsaswellasrepairssingle-strandednicksinduplexDNA.Intheabsenceof20-30%PEG6000,T3DNALigasedisplaysaverylowefficiencyforblunt-endedligation(1).T3DNALigasedisplaysahigherefficiencyforjoiningA/ToverhangsthanC/Gmatchedends(1).T3DNALigaseretains95%ofitsactivityin1.0MNaClorKCl,withanoptimalconcentrationof300mM(1).
Application: Catalyzestheformationofaphosphodiesterbondbetweena5'-phosphateanda3'-hydroxylterminusinduplexDNA.
Source: ArecombinantE.colistraincarryingtheT3DNALigasegene.
Suppliedin: 20mMTris-HCl 300mMNaCl 1mMDTT 0.1mMEDTA 50%Glycerol pH7.5@25°C
Suppliedwith: 2xRapidLigationBuffer
2xRapidLigationBuffer: 132mMTris-HCl 20mMMgCl2 2mMDTT 2mMATP 15%PEG8000 pH7.6 UnitDefinition: OneWeissunitoftheenzymecatalyzestheconversionof1nmolof[32PPi]intoNorit-adsorbableformin20minat37°C. Enzymeactivityisassayedinthefollowingmixture:66mMTris-HCl(pH7.6),6.6mMMgCl2,0.066mMATP,10mMDTT,3.3µM[32PPi]. OneWeissUnitisapproximatelyequivalentto67cohesiveendunits(CEU).Or1000CEU=15WeissUnit. SpecificActivity: 30,000WeissUnit/mg RecommendedStorageCondition:-20°C
Reference: 1.Cai,Liang,etal.(2004)J.Biochem.135,397-403 |