Description
Recognition site and hydrolysis position:
ACTAGT | A↑CTAGT |
TGATCA | TGATC↓A |
Source: Alteromonas haloplanktis SP
Unit definition:
One unit of the enzyme is the amount required to hydrolyze 1 μg of T7 DNA in 1 hour at 37°C in a total reaction volume of 50 μl.
Assayed on:
T7 DNA
Optimal SE-buffer: SE-buffer B + BSA
Enzyme activity (%):
B | G | O | W | Y | ROSE |
100 | 75 | 25 | 25 | 75 | 100 |
Storage conditions: 10 mM Tris-HCl (pH 7.5); 100 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol; Store at -20°C.
Ligations:
After 20-fold overdigestion with enzyme more than 90% of T7 DNA fragments can be ligated and recut.
Non-specific hydrolisis:
No nonspecific activity was detected after incubation of 1 μg of T7 DNA with 40 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 1x reaction mix to a final concentration of 100 μg/ml.
Do not use BSA for long incubation.
References:
Myakisheva, T.V., Belichenko, O.A., Popichenko, D.V., Dedkov, V.S., Degtyarev, S.Kh. Unpublished observations (2000).