BseC I (Cla I isoschizomer)
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BseC I (Cla I isoschizomer) Technical BrochureTechnical Brochure
 

BseCI is a restriction enzyme purified from Bacillus stearothermophilus.

 

Unit substrate: Lambda DNA.

 

Unit calculation assay conditions: 100 mM NaCl, 50 mM Tris-HCl  (pH 7.9 @ 25°C), 10  mM  MgCl2, 1 mM dithiothreitol, 100 μg/ml BSA. Incubate at 55oC.

 

Absence of contaminants: 150 units of BseCI do not produce any unspecific cleavage products after 16 hrs incubation with 1 μg of λ DNA at 55oC. After 100-fold overdigestion with BseCI 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 7.9 @ 25°C), 0.1 mM EDTA, 1 mM dithiothreitol, 0.15% Triton X-100, 200 μg/ml BSA and 50% glycerol. Store at -20oC.

 

Heat inactivation: No.

 

Methylation Sensitivity:

dam methylation: Blocked by overlapping

dcm methylation: Not sensitive

CpG methylation: Blocked

 

Reference: Rina, M., Dialektakis, D., Clark, D., Pagomenou, M. and Bouriotis, V. (1992). Nucleic Acids Res. 20

 

Percent Activity in MINOTECH Buffers
 L SH 
 10 50 100 75-100 50 100
 
 
General reaction mixture:
10U BseCI 1μl
10x H or K buffer*  2μl 
DNA substrate  <1μg
Sterile ultrapure water  Up to 20 μl
Incubate for 15 min at 55oC  
*In the case of H buffer we recommend the addition of BSA to a final concentration of 100 μg/ml. 

 

Reagents supplied: 10x H and 10x K buffer