Recognition site and hydrolysis position:
Source: Acinetobacter calcoaceticus B1
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: SE-buffer K + BSA
Enzyme activity (%):
Storage conditions: 10 mM Tris-HCl (pH 7.5); 50 mM KCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol.
Store at -20°C.
After 10-fold overdigestion with enzyme 95% of the DNA fragments can be ligated and recut.
No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 10 u.a. of enzyme for 16 hours at 37°C.
Reagents Supplied with Enzyme:
Methylation sensitivity: not tested
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.
Do not use BSA for long incubation.
Degtyarev, S.Kh., Dedkov, V.S., Belichenko, O.A., Verchosina, V.A., Kusner, Y.S., Unpublished observations.