Description
Recognition site and hydrolysis position:
CCANNNNNTGG | CCANNNN↑NTGG |
GGTNNNNNACC | GGTN↓NNNNACC |
Source: Acinetobacter calcoaceticus B7
Unit definition:
One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA (dcm-) in 1 hour at 37°C in a total reaction volume of 50 μl.
Assayed on:
Lambda DNA (dcm-)
Optimal SE-buffer: SE-buffer G
Enzyme activity (%):
B | G | O | W | Y | ROSE |
10 | 100 | 25 | 50 | 50 | 100 |
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.
Ligations:
After 5-fold overdigestion with enzyme 95% of the DNA fragments can be ligated and recut.
Non-specific hydrolisis:
No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 5 u.a. of enzyme for 16 hours at 37°C.
Reagents Supplied with Enzyme:
Methylation sensitivity: Blocked by overlapping Dcm methylation (CmCWGG): CCANNNCCTGG or CCAGGNNNTGG.
Notes: High enzyme concentration may result in star activity.
References:
Dedkov, V.S., Repin, V.E., Rechkunova, N.I., Degtyarev, S.Kh., Verhosina, V.A.,Vinogradova, T.P, Izv. Sib. Otd. Akad. Nauk SSSR 1: 35-37 (1990).