Description
Recognition site and hydrolysis position:
CCANNNNNNTGG | CCANNNNN↑NTGG |
GGTNNNNNNACC | GGTN↓NNNNNACC |
Source: Bacillus stearothermophilus X
Unit definition:
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.
Assayed on:
Lambda DNA
Optimal SE-buffer: SE-buffer O
Enzyme activity (%):
B | G | O | W | Y | ROSE |
10 | 10 | 100 | 75 | 25 | 100 |
Storage conditions: 10 mM Tris-HCl (pH 7.5); 100 mM NaCl; 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 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 5 u.a. of enzyme for 16 hours at 37°C.
Reagents Supplied with Enzyme:
Methylation sensitivity: not tested
Notes:
References:
Wise, R., Schildkraut, I. Unpublished observations (1992).