Description
Recognition site and hydrolysis position:
GCNNNNNNNGC | GCNNNNN↑NNGC |
CGNNNNNNNCG | CGNN↓NNNNNCG |
Source: Bacillus stearothermophilus MW
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 Y
Enzyme activity (%):
B | G | O | W | Y | ROSE |
10 | 25 | 25 | 50 | 100 | 40 |
Storage conditions: 10 mM Tris-HCl (pH 7.5); 50 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol.
Storage at -20°C.
Storage at -70°C is recommended for periods longer than 7 days.
Ligations:
After 5-fold overdigestion with enzyme more than 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 10 u.a. of enzyme for 16 hours at 55°C.
Reagents Supplied with Enzyme:
Methylation sensitivity: not tested
Notes: At 37°C activity is 20% from maximum
References:
Nadeev A.N., Dedkov, V.S., Tomilova J.E., Degtyarev, S.K. Unpublished observations (2002).