Technical Data
E0375
EcoRI 10u/ul
5x5000U
Molecular Biology Storage: -20CShipping: Blue Ice
5'-G^A A T T C-3'
3'-C T T A A^G-5'

Source:
E.coli that carries the cloned ecoRIR gene from Escherichia coli RY13

Supplied With:
R1625: Restriction Enzyme Buffer A, 10X: Supplied as a liquid in 33mM Tris-acetate, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA (pH 7.9 at 37C).
R1625-03: Restriction Enzyme Buffer D, 10X: Supplied in 50mM Tris-HCl, pH 7.5, 10mM MgCl2, 100mM NaCl, 0.02% Triton X-100, 0.1mg/ml BSA.

Concentration: 10u/ul.
(See E0375-05 for 50u/ul).

Unit Definition:
One unit is defined as the amount of enzyme required to digest 1ug of lambda DNA in 1 hour at 37C in 50ul of assay buffer.

Incubation Temperature: 37C

Dilution Buffer:
10mM Tris-HCl, pH 7.4 at 25C, 100mM potassium chloride, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol.

Storage Buffer:
Supplied in 10mM potassium phosphate, pH 7.4, 25C, 300mM NaCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 0.15% Triton X-100, 50% glycerol.
Enzyme Properties:
Stability during Prolonged Incubation: A minimum of 0.2u of EcoRI are required for complete digestion of 1ug of lambda DNA in 16 hours at 37C.

Thermal Inactivation: EcoRI is inactivated by incubation at 65C for 20min.

Digestion of Agarose-Embedded DNA: A minimum of 5u of EcoRI is required for complete digestion of 1ug of agarose-embedded lambda DNA in 16 hours.

Compatible Ends: XapI, MunI, TasI

Number of Recognition Sites in DNA:
Lambda: 5
PhiX174: 0
pBR322; 1
pUC57: 1
pUC18/19: 1
pTZ19R/U: 1
M13mp18/19: 1

Quality Control:
Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 80-fold overdigestion (5u/ug lambda DNA x 16 hours) with EcoRI.
Ligation/Recutting Assay:
The ligation and recleavage assay was replaced with LO test after validating experiments showed LO test ability to trace nuclease and phosphatase activities with sensitivity that is higher than L/R by a factor of 100.
Labeled Oligonucleotide Assay: No detectable degradation of a single-stranded and double-stranded labeled oligonucleotide was observed after incubation with 10u of EcoRI for 4 hours.

Blue/White Cloning Assay: The B/W assay was replaced with LO test after validating experiments showed LO test ability to detect nuclease and phosphatase activities with sensitivity that equals to that of B/W test.

Methylation Effects:
Dam, Dcm, EcoBl, EcoKl: never overlaps - no effect,
CpG: may overlap - cleavage impaired.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.