EcoR I

Enzyme name EcoR I
Prototype EcoRI
Known analogs FunII
SKU SE-E057
Turbo version Available
High-concentration version Available
Restriction type Type IIPiType IIP enzymes are classical Type II restriction enzymes with palindromic recognition sequences and symmetric cleavage sites.
Recognition site
5'… G▼AATTC …3'
3'… CTTAA▲G …5'
Source An E.coli strain that carries the cloned EcoR I gene from Escherichia coli
Optimal buffer SE-buffer EcoRI (100 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 50 mM NaCl; 1 mM DTT.)
Optimal temperature 37 °C
Inactivation temperature 65 °C
Buffer activity
BGOWYROSE
507575755050
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
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.
Ligation After 40-fold overdigestion with enzyme about 95% of the DNA fragments can be ligated and recut.
Nonspecific hydrolysis 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.
Methylation sensitivity not tested
Reagents supplied with enzyme 10 X SE-buffer EcoRI, BSA
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
  4. External publication Albertsen, H.M., Le Paslier, D., Abderrahim, H., Dausset, J., Cann, H., Cohen, D. Nucleiv Acids Res. 17: 808 (1989).
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