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Mbo I
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Mbo I Technical BrochureTechnical Brochure
MboI is a restriction enzyme purified from Moraxella bovis (ATCC 10900).

 

Unit substrate: Lambda DNA (dam-).

 

Unit calculation assay conditions: 100 mM KCl, 10 mM Tris-HCl (pH 8.0 @ 25°C), 10 mM MgCl2, 1mM dithiothreitol, 100 μg/ml BSA. Incubate at 37°C.

 

Absence of contaminants: 20 units of Mbo I do not produce any unspecific cleavage products after 16 hrs incubation with 1 μg of λ DNA (dam–) at 37oC. After 10-fold overdigestion with Mbo I, 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: 65oC for 20 minutes..

 

Methylation Sensitivity:

dam methylation: Blocked

dcm methylation: Not sensitive

CpG methylation: Impaired by overlapping

 

Percent Activity in MINOTECH Buffers
 L M  H  SH  A  K 
50-100  50-100  50-100   50 50-100 100 
 
 
General reaction mixture:
10U MboI 1μl
10x UMboI or K buffer*  2μl 
DNA substrate  <1μg
Sterile ultrapure water  Up to 20 μl
Incubate for 15 min at 37oC  
*In the case of UMboI buffer we recommend the addition of BSA to a final concentration of 100 μg/ml. 

 

Reagents supplied: 10x UMboI and 10x K buffer