Description
Recognition site and hydrolysis position:
CTCGTG | C↑TCGTG |
GAGCAC | GAGCA↓C |
Source: Bacillus stearothermophilus 2B
Unit definition:
One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 60°C in a total reaction volume of 50 μl.
Assayed on:
Lambda DNA
Optimal SE-buffer: SE-buffer Y + BSA
Enzyme activity (%):
B | G | O | W | Y | ROSE |
75 | 25 | 10 | 25 | 100 | 100 |
Storage conditions: 10 mM Tris-HCl (pH 7.5); 200 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 50% glycerol. Store at -20°C.
Ligations:
After 10-fold overdigestion with enzyme 95% 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 5 u.a. of enzyme for 16 hours at 60°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:
Abdurashitov, M.A., Belichenko, O.A., Degtyarev, S.K. Unpublished observations (1995).