BamHI is a restriction enzyme purified from Bacillus amyloliquefaciens H.

Unit substrate: Lambda DNA.

Unit calculation assay conditions: 100 mM NaCl, 10 mM Tris-HCl (pH 7.9 @ 25°C), 5 mM MgCl₂, 1 mM dithiothreitol, 100 μg/ml BSA. Incubate at 37^oC.

Absence of contaminants: 100 units of BamHI incubated for 16 hours at 37^oC with 1 μg of λ DNA resulted in a DNA pattern free of detectable nuclease degradation as determined by agarose gel electrophoresis. After 50-fold overdigestion with BamHI, 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 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: Conditions of low ionic strength, high enzyme concentration, glycerol concentration>5% or pH>8.0 may result in star activity.

Reagents supplied: 10x U_BamHI and 10x K buffer