Description: T4 DNA Ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5\'-phosphate and 3\'-hydroxyl termini in duplex DNA or RNA using ATP as a cofactor. Figure. Ligation of λ DNA/Hind III fragments analyzed by agarose gel electrophoresis.Performances of T4 DNA ligase from MCLAB and another supplier were compared within identical reaction mixture at 37°C for 30 minutes. Paralleled negative control was run without ligase. Application:- Cloning of restriction fragments- Joining linkers and adapters to blunt-ended DNASource:A recombinant E. coli strain carrying the cloned T4 DNA Ligase gene. Supplied In:10 mM Tris-HCl50 mM KCl 1 mM DTT0.1 mM EDTA 50% Glycerol pH 7.4 @ 25°CSupplied With:10x T4 DNA Ligase Buffer500mM Tris-HCl100 mM MgCl250 mM DTT10 mM ATPpH 7.6 @ 25°CUnit Definition:One Weiss unit of the enzyme catalyzes the conversion of 1 nmol of [32PPi] into Norit-adsorbable form in 20 min at 37°C. Enzyme activity is assayed in the following mixture: 66 mM Tris-HCl (pH 7.6), 6.6 mM MgCl2, 0.066 mM ATP, 10 mM DTT, 3.3 µM [32PPi]. One Weiss Unit is approximately equivalent to 67 cohesive end units (CEU). Or 1000 CEU = 15 Weiss Unit. Specific Activity: 300U/µg Recommended Storage Condition: -20°C Reference:1. Engler, M.J. and Richardson, C.C. (1982) P.D. Boyer (Eds.), The Enzymes, 5, pp. 3. San Diego: Academic Press. |
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