Recognition site |
CCC↑GGG
GGG↓CCC
|
Source |
E.coli strain that carries the cloned Sma I gene from Serratia marcescens |
Assayed on |
Lambda DNA (HindIII-digest) |
Unit |
One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA (HindIII-digest) in 1 hour at 25°C in a total reaction volume of 50 μl. |
Optimal SE-buffer |
Y (33 mM Tris-acetate (pH 7.9 at 25oC); 10 mM magnesium acetate; 66 mM potassium acetate; 1 mM DTT.) |
Enzyme activity (%) |
B |
G |
O |
W |
Y |
0 - 10 |
0 - 10 |
0 - 10 |
0 - 10 |
100 |
|
Optimal temperature |
25oC
|
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; Store at -20oC. |
Ligations |
After 20-fold overdigestion with enzyme more than 90% of the DNA fragments can be ligated (by using of high concentration T4 DNA Ligase and 10% PEG) and recut. |
Non-specific hydrolisis |
No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 40 u.a. of enzyme for 16 hours at 25oC. |
Reagents Supplied with Enzyme |
10 X SE-buffer Y |
Methylation sensitivity |
not tested |
Inactivation 20 minutes under |
65oC
|