EcoR I

Recognition site:
G^AATTC

Source:
Purified from a strain of E. coli that bears the cloned EcoR I gene from Escherichia coli RY13.

Storage buffer:
50mM Tris-HCl (pH 7.2), 300 mM NaCl, 5 mM EGTA, 0.5 mM EDTA, 5 mM 2-mercaptoethanol, 0.5 mg/ml BSA, 50% (v/v) glycerol, 0.2% (w/v) Triton X-100.

Reaction condition:
REACT 3 Buffer at 37ºC.

Typical ligation/recut assay results:
EcoR I –cleaved λ DNA is ≥95% ligated after 1 h. Of the ligated fragments, 100% are cleaved (recut) by EcoR I.

Recommended storage conditions:
-20 ºC.

Comments:
Does not cleave DNA when either the 3’ A or the C residue is N6-methyladenine or 5-methylcytosine, respectively. Partially resistant to heat inactivation (10 min, 65 ºC). Glycerol concentrations >5% (v/v), low-salt, or high-pH conditions may alter specificity.