EcoR I

Description

Recognition site and hydrolysis position:

Source: An E.coli strain that carries the cloned EcoR I gene from Escherichia coli
Unit definition:
One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 37°C in a total reaction volume of 50 μl.
Assayed on:
Lambda DNA
Optimal SE-buffer: SE-buffer EcoRI
Enzyme activity (%):

Storage conditions: 10 mM Tris-HCl (pH 7.5); 200 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol; Store at -20°C.
Ligations:
After 40-fold overdigestion with enzyme about 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 40 u.a. of enzyme for 16 hours at 37°C. Reagents Supplied with Enzyme 10 X SE-buffer EcoRI, BSA
Reagents Supplied with Enzyme:
Methylation sensitivity: not tested
Notes: High enzyme concentration and using of nonoptimal buffer may result in star activity.
Do not use BSA for long incubation.
To obtain 100% activity, BSA should be added to the 1 x reaction mix to a final concentration of 100 μg/ml.
References:
Albertsen, H.M., Le Paslier, D., Abderrahim, H., Dausset, J., Cann, H., Cohen, D. Nucleiv Acids Res. 17: 808 (1989).
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