Description
Recognition site and hydrolysis position:
GACNNNNNNGTC | GACNNNN↑NNGTC |
CTGNNNNNNCAG | CTGNN↓NNNNCAG |
Source: Deinococcus species D2
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 Y
Enzyme activity (%):
B | G | O | W | Y | ROSE |
75 | 75 | 25 | 50 | 100 | 30 |
Storage conditions: 10 mM Tris-HCl (pH 7.5); 50 mM KCl; 0,1 mM EDTA; 200 μg/ml BSA; 7 mM 2-mercaptoethanol; 50% glycerol; Store at -20°C.
Ligations:
After 10-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 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:
Abdurashitov, M.A., Kileva, E.V., Dedkov, V.S., Degtyarev, S.K. Unpublished observations (1996).