|Source||Acinetobacter calcoaceticus W2131|
|Assayed on||Lambda DNA (dam-)|
|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.|
|Optimal SE-buffer||Y (33 mM Tris-acetate (pH 7.9 at 25°C); 10 mM magnesium acetate; 66 mM potassium acetate; 1 mM DTT.) + BSA|
|Enzyme activity (%)|
|75 - 100||50 - 75||0 - 10||0 - 10||100||30|
|Storage conditions||10 mM Tris-HCl (pH 7.5); 100 mM KCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol; Store at -20°C.|
|Ligation||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.|
|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.
|Reagents Supplied with Enzyme||
10 X SE-buffer Y, BSA|
|Methylation sensitivity||Blocked by Dam methylation (GmATC) GGATC|
|Inactivation 20 minutes under||65oC|
|Notes||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.
|Quality control||Restriction Endonucleases: Quality Control|
|MSDS:||Download MSDS as PDF|
|References:||Abdurashitov, M.A., Belichenko, O.A., Shevchenko, A.V., Degtyarev, S.Kh.
Unpublished observations. (1994)|