Description
Recognition site and hydrolysis position:
| CACNNNGTG | CACNNN↑GTG |
| GTGNNNCAC | GTG↓NNNCAC |
Source: An E.coli strain, that carries the cloned gene Dra III from Deinococcus radiophilus
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 2K
Enzyme activity (%):
| B | G | O | W | Y | ROSE |
| 25 | 50 | 75 | 75 | 50 | 100 |
Storage conditions: 10 mM Tris-HCl (pH 7.5); 300 mM NaCl; 0.1 mM EDTA; 1 mM DTT; 200 μg/ml BSA; 50% glycerol. Store at -20°C.
Ligations:
After 10-fold overdigestion with enzyme 70% of the DNA fragments can be ligated and recut. In the presence of 10%PEG ligation is better.
Non-specific hydrolisis:
No nonspecific activity was detected after incubation of 1 μg of DNA with 5 u.a. of enzyme for 16 hours at 37°C.Reagents Supplied with Enzyme
10 X SE-buffer 2K, BSA
Reagents Supplied with Enzyme:
Methylation sensitivity: not tested
Notes: High enzyme concentration may result in star activity.
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:
de Wit, C.M., Dekker, B.M.M., Neele, A.C., de Waard, A., FEBS Lett. 180: 219-223 (1985)Grosskopf, R., Wolf, W., Kessler, C., Nucleic Acid res. 13: 1517-1528 (1985)
