Unit definition: One unit is defined as amount of enzyme required to produce a complete digest of 1 µg Lambda DNA in 60 min in a total reaction volume of 50µL under the assay conditions.
Unit substrate: Lambda DNA
Unit calculation assay conditions: 100 mM NaCl, 50 mM Tris-HCl (pH 7.9 @ 25°C), 10 mM MgCl2, 1mM DTT, 100 μg/ml BSA. Incubation temperature 37 °C
Absence of contaminants: 50 units of Bgl II do not produce any unspecific cleavage products after 16 hrs incubation with 1µg of Lambda DNA at 37 °C. After 50-fold overdigestion with Bgl II greater than 95% of the DNA fragments can be ligated and recleaved with this enzyme.
Storage buffer: Bgl II is supplied in 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.
Inactivation: Bgl II cannot be heat inactivated in 20min at 80 °C. The enzyme can be removed by spin column purification. It is also possible to stop the digest by addition of EDTA to a final concentration of >10mM or SDS to a concentration of 0.1 %. Alternatively use our environmentally friendly 6x orange gel loading buffer. For further enzymatic reactions remove SDS or EDTA by spin column-based PCR purification kit.
dam methylation: Not sensitive
dcm methylation: Not sensitive
CpG Methylation: Not sensitive
Activity in Simplebiotech buffers:
10x Buffer H: 1000mM NaCl, 500mM Tris-HCl (pH 7.9 at 25 °C), 100mM MgCl2, 1mg/mL BSA
10x Simple buffer: 500 mM Potassium Acetate, 200 mM Tris-acetate (pH 7.9 at 25°C), 100 mM Magnesium Acetate, 1000 μg/ml BSA
Rapid digest within 15 minutes.