Properties & Usage
Unit Definition
One unit is defined as the amount of enzyme required to protect 0.5 µg T4gt-DNA against cleavage by MfeI restriction endonuclease. Reaction Conditions
1X NEBuffer™ 4
Supplement with 40 µM Uridine Diphosphate Glucose
Incubate at 37°C
1X NEBuffer™ 4
50 mM Potassium Acetate
20 mM Tris-acetate
10 mM Magnesium Acetate
1 mM DTT
(pH 7.9 @ 25°C)
Diluent Compatibility
Storage Buffer
20 mM KPO4
200 mM NaCl
0.25 mM DTT
0.1 mM EDTA
50% Glycerol
pH 7 @ 25°C
Heat Inactivation
65°C for 10 minutes Molecular Weight
Theoretical: 40666 kDa
Unit Assay Conditions
0.5 µg T4gt-DNA, 1X NEBuffer 4 and 40 µM UDP-Glucose in a 30 µl reaction. Incubate for 1 hour at 37°C followed by 10 minutes at 65°C. The extent of protection by T4 -BGT is determined by the addition of 20 µl 1X NEBuffer 4 and 10 units of MfeI. Incubation at 37°C for 30 minutes is followed by analysis on agarose gels.
Enzyme Properties
Activity in NEBuffers:
NEBuffer 1 100
NEBuffer 2 50
NEBuffer 3 50
NEBuffer 4 100%
Survival in a Reaction: A minimum of 0.16 unit for 16 hours is required to protect 0.5 µg T4 gt-DNA against cleavage by MfeI.