Description
Recognition site and hydrolysis position:
CCNNNNNNNGG | CCNNNNN↑NNGG |
GGNNNNNNNCC | GGNN↓NNNNNCC |
Source: Bacillus schlegelii 4
Unit definition:
One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 55°C in a total reaction volume of 50 μl.
Assayed on:
Lambda DNA
Optimal SE-buffer: SE-buffer W + BSA
Enzyme activity (%):
B | G | O | W | Y | ROSE |
75 | 75 | 50 | 100 | 25 | 90 |
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 10-fold overdigestion with enzyme more than 90% 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 20 u.a. of enzyme for 16 hours at 55°C.
Reagents Supplied with Enzyme:
Methylation sensitivity: not tested
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.
References:
Repin, V.E., Serov, G.D., Puchkova, L.I., Tereshenko, T.A., Lebedev, L.R., Chigikov, V.E. Bioorg. Khim. 19: 583-585 (1993).