Description
Recognition site and hydrolysis position:
GCAATGNN | GCAATGNN↑ |
CGTTACNN | CGTTAC↓NN |
Source: Bacillus stearothermophilus 3D
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 G
Enzyme activity (%):
B | G | O | W | Y | ROSE |
10 | 100 | 25 | 50 | 75 | 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 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 10 u.a. of enzyme for 16 hours at 60°C.
Reagents Supplied with Enzyme:
Methylation sensitivity: not tested
Notes:
References:
Abdurashitov, M.A., Bondar, T.S., Shevchenko, A.V., Dedkov, V.S., Degtyarev, S.K. Unpublished observations (1996).
Chernukhin V.A, Abdurashitov M.A., Tomilov V.N., Gonchar D.A., Degtyarev S.Kh. Comparative analysis of mouse chromosomal DNA digestion with restriction endonucleases in vitro and in silico // Translated from “Ovchinnikov bulletin of biotechnology and physical and chemical biology” V.3, No 4, pp 19-27, 2007
V.A. Chernukhin, M.A. Abdurashitov, V.N. Tomilov, D.A. Gonchar, S.Kh. Degtyarev Comparative restriction enzymes analysis of rat chromosomal DNA in vitro and in silico // Translated from “Ovchinnikov bulletin of biotechnology and physical and chemical biology” V.2, No 3, pp 39-46, 2006