* Size:

BstE II Technical BrochureTechnical Brochure

BstEII is a restriction enzyme purified from Bacillus stearothermophilus.


Unit substrate: Lambda DNA.


Unit calculation assay conditions: 100 mM KCl, 10 mM Tris-HCl (pH 7.4 @ 25°C), 5 mM MgCl2, 1 mM dithiothreitol, 0.1% Triton X-100, 100 μg/ml BSA. Incubate at 60oC.


Absence of contaminants: 150 units of BstEII do not produce any unspecific cleavage products after 16 hrs incubation with 1 μg of λ DNA at 60oC. After 100-fold overdigestion with BstEII, 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 7.4), 0.1 mM EDTA, 1 mM dithiothreitol, 200 μg/ml BSA and 50% glycerol. Store at -20oC.


Heat inactivation: No.


Methylation Sensitivity:

dam methylation: Not sensitive

dcm methylation: Not sensitive

CpG methylation: Not sensitive


Note: BstE II exhibits 10-15% activity at 37°C.


Percent Activity in MINOTECH Buffers
 L SH 
 50 50-75 75-100 50 75 100
General reaction mixture:
10U BstEII 1μl
10x UBstEII or K buffer*  2μl 
DNA substrate  <1μg
Sterile ultrapure water  Up to 20 μl
Incubate for 15 min at 60oC  
*In the case of UBstEII buffer we recommend the addition of BSA
to a final concentration of 100 μg/ml.


Reagents supplied: 10x UBstEII and 10x K buffer