T3 DNA Ligase

T3 DNA Ligase catalyzes the formation of a phosphodiester bond between a 5′ phosphate and a 3′ hydroxyl terminus in duplex DNA. The enzyme will join blunt ends and cohesive ends as well as repair single stranded nicks in duplex DNA.

Description:
T3 DNA Ligase joins blunt ends and cohesive ends as well as repairs single-stranded nicks in duplex DNA. In the absence of 20-30% PEG 6000, T3 DNA Ligase displays a very low efficiency for blunt-ended ligation. T3 DNA Ligase displays a higher efficiency for joining A/T overhangs than C/G matched ends. T3 DNA Ligase retains 95% of its activity in 1.0 M NaCl or KCl, with an optimal concentration of 300 mM⁽¹⁾.

Application:
Catalyzes the formation of a phosphodiester bond between a 5'-phosphate and a 3'-hydroxyl terminus in duplex DNA.

Source:
A recombinant E. coli strain carrying the T3 DNA Ligase gene.

Supplied in:
20 mM Tris-HCl
300 mM NaCl
1 mM DTT
0.1 mM EDTA
50% Glycerol
pH 7.5 @ 25°C

Supplied with:
2x Rapid Ligation Buffer

2x Rapid Ligation Buffer:
132 mM Tris-HCl
20 mM MgCl₂
2 mM DTT
2 mM ATP
15% PEG 8000
pH 7.6
 

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: 30,000 Weiss Unit/mg

Recommended Storage Condition: -20°C