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'

Supplied With:
R1625: Restriction Enzyme Buffer A, 10X
R1625-03: Restriction Enzyme Buffer D, 10X

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:
For short-term storage (3-4 weeks):
Dilute with Diluent Buffer (R1625-05, 10mM Tris-HCl, pH 7.4 at 25C, 100mM potassium chloride, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol.

For long-term storage:
Dilute with Storage Buffer (10mM potassium phosphate, pH 7.4, 25C, 300mM NaCl, 1mM EDTA, 7mM 2-mercaptoethanol, 0.2mg/ml BSA, 0.15% Triton X-100, 50% glycerol).

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: After 50-fold overdigestion (3u/ug DNA x 17 hours) with EcoRI, more than 90% of the DNA fragments can be ligated at a 5'-termini concentration of 0.02uM. More than 90% of these can be recut.

Labeled Oligonucleotide Assay: No detectable degradation of a single-stranded and double-stranded labeled oligonucleotide was observed after incubation with 10u of restriction endonuclease for 4 hours.

Blue/White Cloning Assay: pUC57 was digested at a unique site with 10u of enzyme for 16 hours. After religation and transformation 0.2% of white colonies were detected.


Methylation Effects: EcoRI cuts neither Gm6AATTC nor GAm6ATTC, but cuts GAATTm5C, GAATThm5C and GAAhm5Uhm5UC.

Storage and Stability:
Store at -20C. Stable for 12-12 months.
Source: E.coli that carries the cloned ecoRIR gene from Escherichia coli RY13

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

Enzyme Properties:
Star Activity: Star Activity appears at 10-fold overdigestion (10u x 1 hour). A large excess of the enzyme (10u/ug DNA x 16 hours), low salt concentration, high pH, or the replacement of Mg2+ by Mn2+ results in star activity.

Stability during Prolonged Incubation: A minimum of 0.2u of enzyme are required for complete digestion of 1ug of lambda DNA in 16 hours at 37C.

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

Digestion of Agarose-Embedded DNA: A minimum of 5u of enzyme 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

Important Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.