EcoR I
* Size:

EcoR I Technical BrochureTechnical Brochure

EcoRI is a restriction enzyme purified from E. coli RY 13.


Unit substrate: Lambda DNA.


Unit calculation assay conditions: 50 mM NaCl, 100 mM Tris-HCl (pH 7.4 @ 25oC), 5 mM MgCl2, 0.025% Triton X-100, 100 μg/ml bovine serum albumin and DNA. Incubate at 37oC.


Absence of contaminants: 100 units of EcoR I do not produce any unspecific cleavage products after 16 hrs incubation with 1 μg of Lambda DNA at 37oC. After 50-fold overdigestion with EcoR I greater than 98% of the DNA fragments can be ligated and recut with this enzyme.


Storage buffer: 300 mM NaCl, 5 mM KPO4, (pH 7.4), 0.1 mM EDTA, 1 mM dithiothreitol, 0.15% Triton X-100, 200 μg/ml bovine serum albumin and 50% glycerol. Store at -20oC.


Heat inactivation: 65oC for 20 minutes.


Methylation Sensitivity:

dam methylation: Not sensitive

dcm methylation: Not sensitive

CpG methylation: Impaired by overlapping

Star activity: Conditions of low ionic strength, high enzyme concentration, glycerol concentration>5%, or pH>8.0 may result in star activity.


Percent Activity in MINOTECH Buffers
 L SH 
25-50  50-75  75  50-75  75  100 
General reaction mixture:  
10U EcoRI 1μl
10x UEcoRI or K buffer * 2μl 
DNA substrate  <1μg
Sterile ultrapure water  Up to 20 μl
Incubate for 15 min at 37oC  
*In the case of UEcoRI buffer we recommend the addition of BSA to a final concentration of 100 μg/ml. 


Reagents supplied: 10x UEcoRI and 10x K buffer