Asu II is a restriction enzyme purified from an isolated strain (#94S).

Unit substrate: Lambda DNA (Hind III digest).

Unit calculation assay conditions: 50 mM NaCl, 10 mM Tris-HCl (pH 7.9 @ 25°C), 10 mM MgCl₂, 1 mM dithiothreitol, 0.1% Triton X-100, 100 μg/ml BSA. Incubate at 37^oC.

Absence of contaminants:  100 units of AsuII do not produce any unspecific  cleavage products after 16 hrs incubation with 1 μg of λ DNA/Hind III digest at 37^oC.  After 50-fold overdigestion with AsuII, greater than 95% of the DNA fragments can be ligated and recut with this enzyme.

Storage buffer: 100 mM KCl, 10 mM Tris-HCl (pH 7.9@ 25°C), 0.1 mM EDTA, 1 mM dithiothreitol, 0.15% Triton X-100, 200 μg /ml BSA and 50% glycerol. Store at -20^oC.

Heat inactivation: 65^oC for 20 minutes.

Methylation Sensitivity:

dam methylation: Not sensitive

dcm methylation: Not sensitive

CpG methylation: Blocked

Reagents supplied: 10x U_AsuII and 10x K buffer