Recognition site |
C↑TCGAG
GAGCT↓C
|
Source |
Streptomyces fradiae 274 |
Assayed on |
Lambda DNA (Hind III-digest) |
Unit |
One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA (Hind III-digest) in 1 hour at 50°C in a total reaction volume of 50 μl. |
Optimal SE-buffer |
B (10 mM Tris-HCl (pH 7.6 at 25oC); 10 mM MgCl2; 1 mM DTT.) |
Enzyme activity (%) |
B |
G |
O |
W |
Y |
100 |
75 - 100 |
50 - 75 |
50 - 75 |
75 - 100 |
|
Optimal temperature |
50oC
|
Storage conditions |
10 mM Tris-HCl (pH 7.5); 50 mM KCl; 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 and recut. |
Non-specific hydrolisis |
No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 20 u.a. of enzyme for 16 hours at 50oC. |
Reagents Supplied with Enzyme |
10 X SE-buffer B |
Methylation sensitivity |
Blocked by CTCGmAG methylation.
Not blocked by CTmCGAG methylation. |
Inactivation 20 minutes under |
65oC
|