Description
Recognition site and hydrolysis position:
TGGCCA | TGG↑CCA |
ACCGGT | ACC↓GGT |
Source: Microbacterium oxydans
Unit definition:
One unit of the enzyme is the amount required to hydrolyze 1 μg of Lanbda DNA in 1 hour at 37°C in a total reaction volume of 50 μl.
Assayed on:
Lambda DNA (dcm-)
Optimal SE-buffer: SE-buffer O
Enzyme activity (%):
B | G | O | W | Y | ROSE |
10 | 25 | 100 | 75 | 25 | 75 |
Storage conditions: 10 mM Tris-HCl (pH 7.5); 200 mM KCl; 0,1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol; Store at -20°C.
Ligations:
After 20-fold overdigestion with enzyme 80% of the DNA fragments can be ligated and recut. More complete ligation can be reached by using 10% PEG
Non-specific hydrolisis:
No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 40 u.a. of enzyme for 16 hours at 37°C.
Reagents Supplied with Enzyme:
Methylation sensitivity: Cleaved of DNA is impaired by overlapping Dcm methylation(CmCWGG): TGGCCAGG.
Notes:
References:
Chernov, A.P., Belichenko, O.A., Rechkunova, N.I., Andreeva, I.S., Repin, V.E., Degtyarev, S.K. Russian Patent Office (1993). SU 1806191 A3
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