Description
Recognition site and hydrolysis position:
CGGCCG | C↑GGCCG |
GCCGGC | GCCGG↓C |
Source: Bacillus stearothermophilus X3
Unit definition:
One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 50°C in a total reaction volume of 50 μl.
Assayed on:
Lambda DNA
Optimal SE-buffer: SE-buffer O
Enzyme activity (%):
B | G | O | W | Y | ROSE |
10 | 25 | 100 | 50 | 10 | 25 |
Storage conditions: 10 mM Tris-HCl (pH 8.2); 100 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol.
Store at -20°C. Storage at -70 °C is recommended for periods longer than 3 months
Ligations:
After 5-fold overdigestion with enzyme more than 90% of 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 10 u.a. of enzyme for 16 hours at 50°C.
Reagents Supplied with Enzyme:
Methylation sensitivity: Blocked by CG methylation.
Notes:
References:
Kileva, E.V., Dedkov, V.S., Shevchenko, A.V., Abdurashitov, M.A., Degtyarev, S.K. Unpublished observations (1996).