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SseBI (Stu I isoschizomer)
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SseBI

(Stu I isoschizomer)

 Technical Brochure Technical Brochure

SseBI is a restriction enzyme purified from Streptomyces species.

 

Unit substrate: Lambda DNA (HindIII digest).

 

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 and DNA. Incubate at 37oC.

 

Absence of contaminants: 150 units of SseBI do not produce any unspecific cleavage products after 16 hrs incubation with 1 μg of λ DNA/Hind III digest at 37oC. After 50-fold overdigestion with SseBI, 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 7.9 @ 25°C), 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: Not sensitive

dcm methylation: Blocked by overlapping

CpG methylation: Not sensitive

 

Reference: Rina, M., Tzanodaskalaki, M., Karagouni, A., Pagomenou, M. and Bouriotis, V. (1992) Nucleic Acids Res. 20, 1808. 

 

Percent Activity in MINOTECH Buffers
 L M  H  SH  A  K 
50-75  75-100  100  50-75  50  100 
 
 
General reaction mixture:
10U SseBI 1μl
10x H or K buffer*  2μl 
DNA substrate  <1μg
Sterile ultrapure water  Up to 20 μl
Incubate for 15 min at 37oC  
*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