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Specifications

5'-C A C N N N^G T G-3'
3'-G T G^N N N C A C-5'
Catalog #	A0859-75
Concentration	10u/ul
	Source:
	Alcaligenes denitrificans SS 3–028
Buffers Supplied	BR1625-02: Restriction Enzyme Buffer C, 10X and R1625: Restriction Enzyme Buffer A, 10X
	Buffer:
	Supplied in 10mM Tris-HCl (pH 7.5), 10mM MgCl2, 50mM NaCl and 0.1mg/ml BSA. Incubate at 37°C.
Diluent Buffer	Supplied in 10mM Tris-HCl (pH 7.4 at 25°C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol.
Storage Buffer	Supplied in 10mM Tris-HCl (pH 7.5 at 25°C), 100mM KCl, 1mM DTT, 0.1mM EDTA, 0.2mg/ml BSA and 50% glycerol.
Restriction Enzyme Buffer A, 10X	Supplied in 33mM Tris-acetate (pH 7.9 at 37ºC),10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA.
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.
Storage and Stability	Aliquot to avoid repeated freezing and thawing. Store at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Overdigestion Assay
No detectable change in the specific fragmentation pattern is observed after 15-fold overdigestion (15u/ug lambda DNA x 1hour) with AdeI (see Star Activity).
Ligation/Recutting Assay
After 10-fold overdigestion (5u/ug DNA x 2 hours) with AdeI, more than 80% of the DNA fragments can be ligated at a 5'-termini concentration of 0.1uM. 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
The mix of pUC57/HindIII, pUC57/Eco32I and pUC57/PstI digests was incubated with 10 units of enzyme for 16 hours. After religation and transformation 0.2% of white colonies were detected.
Star Activity
A large excess of enzyme (20u/µg DNA x 1 hour) may result in star activity.
Methylation Effect
DAM, DCM	Never overlaps - no effect,
EcoKl, EcoBl	May overlap- effect not determined, CpG: may overlap - cleavage impaired.