MCLAB/T4 DNA Ligase/TL300/5000 Weiss units

Description:
T4DNALigasecatalyzestheformationofaphosphodiesterbondbetweenjuxtaposed5'-phosphateand3'-hydroxylterminiinduplexDNAorRNAusingATPasacofactor.

Figure.LigationofλDNA/HindIIIfragmentsanalyzedbyagarosegelelectrophoresis.
PerformancesofT4DNAligasefromMCLABandanothersupplierwerecomparedwithinidenticalreactionmixtureat37°Cfor30minutes.Parallelednegativecontrolwasrunwithoutligase.

Application:
-Cloningofrestrictionfragments
-Joininglinkersandadapterstoblunt-endedDNA

Source:
ArecombinantE.colistraincarryingtheclonedT4DNALigasegene.
 

SuppliedIn:
10mMTris-HCl
50mMKCl 
1mMDTT
0.1mMEDTA 
50%Glycerol 
pH7.4@25°C

SuppliedWith:
10xT4DNALigaseBuffer
500mMTris-HCl
100mMMgCl₂
50mMDTT
10mMATP
pH7.6@25°C

UnitDefinition:
OneWeissunitoftheenzymecatalyzestheconversionof1nmolof[³²PP_i]intoNorit-adsorbableformin20minat37°C. Enzymeactivityisassayedinthefollowingmixture:66mMTris-HCl(pH7.6),6.6mMMgCl₂,0.066mMATP,10mMDTT,3.3µM[³²PP_i].

OneWeissUnitisapproximatelyequivalentto67cohesiveendunits(CEU).Or1000CEU=15WeissUnit.

SpecificActivity:300U/µg

RecommendedStorageCondition: -20°C

Reference:
1.Engler,M.J.andRichardson,C.C.(1982)P.D.Boyer(Eds.),TheEnzymes,5,pp.3.SanDiego:AcademicPress.