EcoICR I

Description

Recognition site and hydrolysis position:

Source: Escherichia coli ICR
Unit definition:
One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA (HindIII-digest) in 1 hour at 37°C in a total reaction volume of 50 μl.
Assayed on:
Lambda DNA (HindIII-digest)
Optimal SE-buffer: SE-buffer G
Enzyme activity (%):

Storage conditions: 10 mM Tris-HCl (pH 7.5); 50 mM KCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 50% glycerol; Store at -20°C.
Ligations:
After 5-fold overdigestion with enzyme > 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 10 u.a. of enzyme for 16 hours at 37°C. Reagents Supplied with Enzyme 10 X SE-buffer G, BSA
Reagents Supplied with Enzyme:
Methylation sensitivity: not tested
Notes: To obtain 100% activity, BSA should be added to the 1 x reaction mix to a final concentration of 100 μg/ml
Do not use BSA for long incubation.
References:
Sugisaki H., Maekawa Y., Kanazawa S., Takanami M. Bull. Inst. Chem. Res. Kyoto Univ. 60: 328-335 (1982).Yankovsky N.K., Fonstein M.Yu., Lashina S.Yu., Bukanov N.O., Yakubovich N.V., Ermakova L.M., Rebentish B.A., Janulaitis A., Debabov V.G. Gene 81: 203-210 (1989).