Recognition Sequence :
Unit Definition :
One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA (dam-) in 1 hour at 37°C in a total reaction volume of 50 μl.
Reaction Temperature :
Storage Buffer :
10 mM Tris-HCl (pH 7.5); 100 mM KCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol
After 3-fold overdigestion with enzyme about 50% of the DNA fragments can be ligated with T4 DNA ligase and recut. In the presence of 10% PEG ligation is better.
Acinetobacter calcoaceticus W2131
Assayed on :
Lambda DNA (dam-)
Working buffer :
Y (33 mM Tris-acetate (pH 7.9 at 25°C); 10 mM magnesium acetate; 66 mM potassium acetate; 1 mM DTT.) + BSA
Non-specific hydrolisis :
No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 6 u.a. of enzyme for 16 hours at 37°C.
Concentration, u.a./ml :
20min Under 65°C
Methylation sensitivity :
Blocked by Dam methylation (GmATC) GGATC
Reagents Supplied :
10 X SE-buffer Y, BSA
To obtain 100% activity, BSA should be added to the 1x reaction mix to a final concentration of 100 μg/ml.
Do not use BSA for long incubation.