T4 DNA Ligase

Description

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.

Applications:

• Cloning of restriction fragments
• Joining linkers and adapters to blunt-ended DNA

Source:
A recombinant E. coli strain carrying the cloned T4 DNA Ligase gene.

Supplied In:
10 mM Tris-HCl
50 mM KCl
1 mM DTT
0.1 mM EDTA
50% Glycerol
pH 7.4 @ 25°C

Supplied With:
10x T4 DNA Ligase Buffer

10x T4 DNA Ligase Buffer:
500mM Tris-HCl
100 mM MgCl₂
50 mM DTT
10 mM ATP
pH 7.6 @ 25°C

Unit Definition:
One Weiss unit of the enzyme catalyzes the conversion of 1 nmol of [³²PP_i] 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 [³²PP_i].

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.

Additional information

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