Technical Data
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5'-G G T C T C(N)1^-3'
3'-C C A G A G(N)5^-5'


E. coli that carries the cloned eco31IR gene from E. coli RFL31

Reaction Buffer (10X) (E0373-55A):
Tris-HCl (pH 7.5), 10mM MgCl2, 50mM NaCl and 0.1mg/ml BSA. Incubate at 37C

Restriction Enzyme Buffer A, 10X:
33mM Tris-acetate (pH 7.9), 10mM magnesium acetate, 66mM potassium acetate and 0.1mg/ml BSA. Incubate at 37C. Supplied to simplify buffer selection for double digests.

Diluent Buffer:
10mM Tris-HCl (pH 7.4 at 25C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol, or Storage Buffer.

Storage Buffer:
10mM Tris-HCl (pH 7.5 at 25C), 200mM KCl, 1mM DTT, 0.1mM EDTA, 0.2mg/ml BSA and 50% glycerol.
Overdigestion Assay:
No detectable change in the specific fragmentation pattern is observed after 80-fold overdigestion (5u/ug lambda DNA x 16 hours) with Eco31I (see Star Activity).

Ligation/Recutting Assay:
After 50-fold overdigestion (3u/ug DNA x 17 hours) with Eco31I, more than 95% of the DNA fragments can be ligated at a 5'-termini concentration of 0.05uM. More than 95% of these can be recut.

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

Star Activity:
A large excess of enzyme (10u/ug DNA x 16 hours) may result in star activity.

Methylation Effects:
Sequence GAGACm5C is cleaved by Eco31I at least 50-fold slower than an unmethylated site. Overlapping Dcm or CG methylation may influence DNA cleavage.

Stability during Prolonged Incubation:
A minimum of 0.3 units of enzyme is 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 5 units of enzyme is required for digestion of 1ug of agarose-embedded lambda DNA in 16 hours.

Number of Recognition Sites in DNA:
Lambda: 2
PhiX174: 0
M13mp18/19: 0
pBR322: 1
pUC18/19: 1
pUC57: 1
pTZ19R/U: 1
pBluescriptIIKS(-/+): 1
pBluescriptIISK(-/+): 1
pACYC177: 1
pACYC184: 0

Unit Definition:
One unit is defined as the amount of enzyme required to digest 1ug of lambda DNA dcm--HindIII fragments in 1 hour at 37C in 50ul of assay buffer. Assayed using lambda DNA dcm-, as one of two Eco31I recognition sites is difficult to cleave.

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.