Unit substrate: Lambda DNA.

Unit calculation assay conditions: 100 mM KCl, 10 mM Bis-Tris-Propane (pH 7.0 @ 25°C), 10 mM MgCl₂, 100 μg/ml BSA. Incubate at 37°C.

Absence of contaminants: 600 units of Sca I incubated for 16 hours at 37ºC with 1 μg of λ DNA resulted in a DNA pattern free of detectable nuclease degradation as determined by agarose gel electrophoresis. After 100-fold overdigestion with Sca I, greater than 90% of the DNA fragments can be ligated and recut with this enzyme.

Storage buffer: 50 mM KCl, 10 mM Tris-HCl  (pH 7.4), 0.1 mM EDTA, 1 mM dithiothreitol, 500 μg/ml BSA and 50% glycerol. Store at -20^oC.

Heat inactivation: 80^oC for 20 minutes.

Methylation Sensitivity:

dam methylation: Not sensitive

dcm methylation: Not sensitive

CpG methylation: Not sensitive

Star activity: Large excess of the enzyme may results in the appearance of star activity.

Reagents supplied: 10x U_ScaI buffer