Description
Recognition site and hydrolysis position:
GTCTCN | GTCTCN↑ |
CAGAG(N)5 | CAGAG(N)5↓ |
Source: Bacillus stearothermophilus MA
Unit definition:
One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 55°C in a total reaction volume of 50 μl.
Assayed on:
Lambda DNA
Optimal SE-buffer: SE-buffer W + BSA
Enzyme activity (%):
B | G | O | W | Y | ROSE |
25 | 50 | 50 | 100 | 75 | 100 |
Storage conditions: 10 mM Tris-HCl (pH 7.5); 100 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 100 μg/ml BSA; 50% glycerol; Store at -20°C.
Ligations:
After 50-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 100 u.a. of enzyme for 16 hours at 55°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:
Chernukhin, V.A., Mikhnenkova, N.A., Kileva, E.V., Mezentseva, N.V., Dedkov, V.S., Degtyarev, S.K. Unpublished observations (2005)
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