Technical Data
Ecl136II (EcoICRI)
Molecular Biology Storage: -20CShipping: Blue Ice
5'.....G A G^C T C.....3'
3'.....C T C^G A G.....5'

Unlike SacI, Ecl136II produces DNA fragments with blunt ends.

Source: Enterobacter cloacae RFL136

Concentration: 10 units/ul

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

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).

Supplied with:
R1625-55: Restriction Enzyme Buffer for Ecl136II, 10X: Supplied as a liquid 10mM Bis-Tris Propane-HCl, pH 6.5, 10mM MgCl2, 0.1mg/ml BSA. Incubate at 37C.

Diluent and Storage Buffer:
10mM Tris-HCl , pH 7.4 at 25C, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol.
Quality Control:
Overdigestion Assay:
No detectable change in the specific fragmentation pattern is observed after 320-fold overdigestion (20u/ug lambda DNA x 16 hours) with Ecl136II.

Ligation/Recutting Assay :
After 50-fold overdigestion (3u/ug DNA x 17 hours) with Ecl136II, more than 90% of the DNA fragments can be ligated at a 5'-termini concentration of 0.01uM. More than 90% 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.

Blue/White Cloning Assay:
pUC57 was digested at a unique site within lacZ gene with 10 units of enzyme for 16 hours. After religation and transformation 1% of white colonies were detected.

Methylation Effects:
The hemi-methylated sequence GAGCTm5C is cleaved by Ecl136II at a significantly slower rate. Cleavage is blocked when both strands are methylated at positions: 5' GAGCTm5C3' 3'm5CTCGA G5' [Overlapping CG methylation may influence DNA cleavage]

Stability during Prolonged Incubation:
A minimum of 0.2 units 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.

Number of Recognition Sites in DNA:
Lambda 2
PhiX174 0
M13mp18/19 1
pBR322 0
pUC18/19 1
pUC57 1
pTZ19R/U 1

Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.