BamHI
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BamH I
Technical BrochureTechnical Brochure
 

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 37oC.

 

Absence of contaminants: 100 units of BamHI 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 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 -20oC.

 

Heat inactivation: 80oC 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.

Percent Activity in MINOTECH Buffers
 L SH 
 75 75-100 100 50-75 75 100
 

 

General reaction mixture:
10U BamH I
1μl
10x UBamHI or K buffer * 2μl 
DNA substrate 

<1μg

Sterile ultrapure water  Up to 20 μl
Incubate for 15 min at 37oC  

*In the case of UBamHI buffer we recommend the addition
of BSA to a final concentration of 100 μg/ml.

Reagents supplied: 10x UBamHI and 10x K buffer