|Source||An. E.coli strain that carries the cloned Acu I gene from Acinetobacter calcoaceticus SRW4|
|Assayed on||Lambda DNA|
|Unit definition||One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA 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 + SAM|
|Enzyme activity (%)|
|25 - 50||50 - 75||50 - 75||75 - 100||100||50|
|Storage conditions||10 mM Tris-HCl (pH 7.5); 100 mM NaCl; 0.1 mM EDTA; 1 mM DTT; 200 μg/ml BSA; 50% glycerol; Store at -20°C.|
|Ligation||After 2-fold overdigestion with enzyme about 80% of the DNA fragments can be ligated. Of these, 80% can be recut.|
|Non-specific hydrolisis||No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 1 u.a. of enzyme for 16 hours at 37°C.
|Reagents Supplied with Enzyme||
10 X SE-buffer Y, BSA, SAM|
|Methylation sensitivity||not tested|
|Inactivation 20 minutes under||65oC|
|Notes||High enzyme concentration may result in star activity.|
To obtain 100% activity, BSA should be added to the 1x reaction mix to a final concentration of 100 μg/ml and SAM should be added to a final concentration 10 μM. It is possible to use a final SAM concentration of 10-64 μM. In this case 32 mM SAM solution should be diluted in 3200-500 times, correspondingly. For the small volume of reaction mix (10-20 μl) the stock SAM solution may be previously diluted up to 1 mM (in 32 times) with 5 mM solution of H2SO4 or sterile water and stored at -20°C.
Do not use BSA for long incubation.
|Quality control||Restriction Endonucleases: Quality Control|
|MSDS:||Download MSDS as PDF|
|References:||Degtyarev, S.Kh., Kileva, E.V., Dedkov, V.S. Unpublished observations (2001). |