Description
Recognition site and hydrolysis position:
GGCGCC | GGC↑GCC |
CCGCGG | CCG↓CGG |
Source: Enterobacter gergoviae
Unit definition:
One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA (HindIII-digest) in 1 hour at 37°C in a total reaction volume of 50 μl.
Assayed on:
Lambda DNA (HindIII-digest)
Optimal SE-buffer: SE-buffer B
Enzyme activity (%):
B | G | O | W | Y | ROSE |
100 | 75 | 10 | 50 | 75 | 100 |
Storage conditions: 10 mM Tris-HCl (pH 7.5); 250 mM NaCl; 0,1 mM EDTA; 7 mM 2-mercaptoethanol; 50% glycero; Store at -20°C.
Ligations:
After 5-fold overdigestion with enzyme 70% 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 37°C.
Reagents Supplied with Enzyme:
Methylation sensitivity: not tested
Notes: To obtain 100% activity, BSA should be added to the 1 x reaction mix to a final concentration of 100 μg/ml.
Do not use BSA for long incubation.
References:
Dedkov, V.S., Myakisheva, T.V., Kileva, E.V., Degtyarev, S.K. Unpublished observations (1996).