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BclI
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Bcl I Technical BrochureTechnical Brochure
 

BclI is a restriction enzyme purified from Bacillus caldolyticus.

 

Unit substrate: Lambda DNA (dam-).

 

Unit calculation assay conditions: 50 mM NaCl, 10 mM Tris-HCl (pH 7.9 @ 25°C), 10 mM MgCl2, 1 mM dithiothreitol, 100 μg/ml BSA. Incubate at 50oC.

 

Absence of contaminants: 100 units of BclI do no produce any unspecific clevage products after 16 hrs incubation with 1 μg of λ DNA (dam-) at 50°C. After 50-fold overdigestion with BclI, 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: No.

 

Methylation Sensitivity:

dam methylation: Blocked

dcm methylation: Not sensitive

CpG methylation: Not sensitive

 

Percent Activity in MINOTECH Buffers
 L M  H  SH  A  K 
10-25 100 75 50-75 10-25 100
 

 

General reaction mixture:
10U BclI 1μl
10x M or K buffer * 2μl 
DNA substrate  <1μg
Sterile ultrapure water  Up to 20 μl
Incubate for 15 min at 50oC  
 

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

Reagents supplied: 10x M and 10x K buffer