Description
Recognition site and hydrolysis position:
GCANNNNNTGC | GCANNNN↑NTGC |
CGTNNNNNACG | CGTN↓NNNNACG |
Source: Bacillus stearothermophilus AP
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 W
Enzyme activity (%):
B | G | O | W | Y | ROSE |
25 | 25 | 75 | 100 | 25 | 100 |
Storage conditions: 10 mM Tris-HCl (pH 7.5); 50 mM KCl; 0,1 mM EDTA; 1 mM DTT; 200 μg/ml BSA; and 50% glycerol Store at -20°C.
Ligations:
After 5-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 5 u.a. of enzyme for 16 hours at 60°C.
Reagents Supplied with Enzyme:
Methylation sensitivity: not tested
Notes: High enzyme concentration may result in star activity. At 37°C activiti is 50% from maximum.
The minimum number of units that resulted in complete digestion of 1 μg of substrate DNA in 16 hours is 0,5.
References:
Abdurashitov, M.A., Belichenko, O.A., Shevchenko, A.V., Dedkov, V.S., Degtyarev, S.Kh. Nucleic Acids Res. 25: 2301-2302 (1997).