Description
Recognition site and hydrolysis position:
AGTACT | AGT↑ACT |
TCATGA | TCA↓TGA |
Source: Zoogloea ramigera SCA
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 |
50 | 25 | 0 | 0 | 100 | 50 |
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 -20°C.
Ligations:
After 20-fold overdigestion with enzyme more than 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 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 1 x reaction mix to a final concentration of 100 μg/ml.
Do not use BSA for long incubation.
References:
Nadeev, A.N., Kileva, E.V., Popichenko, D.V., Dedkov, V.S., Degtyarev, S.K. Unpublished observations.(2003)