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SspI
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SspI Technical Brochure Technical Brochure
SspI is a restriction enzyme purified from Sphaerotilus species.

 

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 37oC.

 

Absence of contaminants: 30 units of SspI do not produce any unspecific cleavage products after 16 hrs incubation with 1 μg of λ DNA at 37°C. After 10-fold overdigestion with SspI, 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 8.0), 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: 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 M  H  SH  A  K 
 10-25 50-75 100  75-100  50  100 
 
 
General reaction mixture:
10U SspI 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