Description
Recognition site and hydrolysis position:
CCNGG | ↑CCNGG |
GGNCC | GGNCC↓ |
Source: Bacillus stearothermophilus SC
Unit definition:
One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA (dcm-) in 1 hour at 55°C in a total reaction volume of 50 μl.
Assayed on:
Lambda DNA (dcm-)
Optimal SE-buffer: SE-buffer Y
Enzyme activity (%):
B | G | O | W | Y | ROSE |
50 | 50 | 50 | 50 | 100 | 60 |
Storage conditions: 10 mM Tris-HCl (pH 7.5); 50 mM KCl; 0,1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; and 50% glycerol. Store at -20°C.
Ligations:
After 5-fold overdigestion with enzyme more than 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 3 u.a. of enzyme for 16 hours at 55°C.
Reagents Supplied with Enzyme:
Methylation sensitivity: Blocked by Dcm-methylation (CmCWGG) CCWGG.
Notes: At 37°C activity is 10% from maximum.
References:
Dedkov, V.S., Muradov, S.V., Kileva, E.V., Popichenko, D.V., Degtyarev, S.K. Unpublished observations (2001).
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
Murat A Abdurashitov, Victor N Tomilov, Valery A Chernukhin, S. Kh. Degtyarev A physical map of human Alu repeats cleavage by restriction endonucleases // BMC Genomics 2008, 9:305
Abdurashitov M.A., Tomilov V.N., Chernukhin V.A., Gonchar D. A., Degtyarev S. Kh. Comparative analysis of human chromosomal DNA digestion with restriction endonucleases in vitro and in silico. // Translated from “Medical genetics” V.6, No 8, pp 29-36, 2007