Unit substrate: Lambda DNA.

Unit calculation assay conditions: 50 mM NaCl, 10 mM Tris-HCl (pH 7.9 @ 25^oC), 10 mM MgCl₂, 1 mM dithiothreitol, 100 μg/ml bovine serum albumin and DNA. Incubate at 37^oC.

Absence of contaminants: 100 units of EcoR V do not produce any unspecific cleavage products after 16 hrs incubation with 1 μg of Lambda DNA at 37^oC. After 20-fold overdigestion with EcoR V, greater than 95% 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, 200 μg/ml bovine serum albumin 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: Blocked by overlapping

Star activity: Conditions of low ionic strength, high enzyme concentration, glycerol concentration>5%, or pH>8.0 may result in star activity.