Forgot your password?
New User?
Remember me
banner banner

You are here:Home » Molecular Biology » MB-Enzymes, Restriction » EcoRI 10u/ul

EcoRI 10u/ul


  For pricing information, USA customers sign in.
  Outside USA? Please contact your distributor for pricing.


5'-G^A A T T C-3'
3'-C T T A A^G-5'
Catalog #E0375
Supplied WithR1625: 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 37°C in 50ul of assay buffer.
Incubation Temperature 37°C
DilutionFor short-term storage (3-4 weeks):
Dilute with Diluent Buffer (R1625-05, 10mM Tris-HCl, pH 7.4 at 25°C, 100mM potassium chloride, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol.
For long-term storageDilute with Storage Buffer (10mM potassium phosphate, pH 7.4, 25°C, 300mM NaCl, 1mM EDTA, 7mM 2-mercaptoethanol, 0.2mg/ml BSA, 0.15% Triton X-100, 50% glycerol).
Quality ControlOverdigestion 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 StabilityStore at -20C. Stable for 12-12 months.
SourceE.coli that carries the cloned ecoRIR gene from Escherichia coli RY13
(See E0375-05 for 50u/ul).
Enzyme Properties
Star ActivityStar 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 IncubationA minimum of 0.2u of enzyme are required for complete digestion of 1ug of lambda DNA in 16 hours at 37°C.
Thermal InactivationEnzyme is inactivated by incubation at 65°C for 20min.
Digestion of Agarose-Embedded DNAA minimum of 5u of enzyme is required for complete digestion of 1ug of agarose-embedded lambda DNA in 16 hours.
Compatible EndsXapI, MunI, TasI
Number of Recognition Sites in DNA
pBR322; 1
Important NoteThis 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.

External Links