Recognition site |
GAT↑C
C↓TAG
|
Source |
Bacillus stearothermophilus KT |
Assayed on |
Lambda DNA (dam-) |
Unit |
One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA (dam-) in 1 hour at 37°C in a total reaction volume of 50 μl. |
Optimal SE-buffer |
W (10 mM Tris-HCl (pH 8.5 at 25oC); 10 mM MgCl2; 100 mM NaCl; 1 mM DTT.) |
Enzyme activity (%) |
B |
G |
O |
W |
Y |
25 - 50 |
50 - 75 |
75 - 100 |
100 |
50 - 75 |
|
Optimal temperature |
37oC
|
Storage conditions |
10 mM Tris-HCl (pH 7.5); 100 mM NaCl; 0,1 mM EDTA; 7 mM 2-mercaptoethanol; 100 μg/ml BSA; 50% glycerol. Store at -20oC. |
Ligations |
After 5-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 5 u.a. of enzyme for 16 hours at 37oC. |
Reagents Supplied with Enzyme |
10 X SE-buffer W |
Methylation sensitivity |
Blocked by overlapping Dam methylation (GmATC) GATC .
Not blocked by CG methylation.
Cut hemimethylated site: 5? GmATC-3?/ 5?GATC-3?/td> |
Inactivation 20 minutes under |
65oC
|