Back to Products
Sfi I
* Size:



Sfi I Technical Brochure Technical Brochure
SfiI is a restriction enzyme purified from Streptomyces fimbriatus (ATCC 15051).

 

Unit substrate: Adenovirus-2 DNA.

 

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 50°C.

 

Absence of contaminants: 100 units of Sfi I do not produce any unspecific cleavage products after 16 hrs incubation with 1 μg of Adeno-2 DNA at 50oC. After 50-fold overdigestion with Sfi I, greater than 95% of the DNA fragments can be ligated and recut with this enzyme.

 

Storage buffer: 300 mM NaCl, 5 mM KPO4  (pH 7.4), 0.1 mM  EDTA, 1 mM dithiothreitol, 0.15% Triton X-100, 500 μg/ml BSA and 50% glycerol. Store at -20oC.

 

Heat inactivation: No.

 

Methylation Sensitivity:

dam methylation: Not sensitive

dcm methylation: Impaired by overlapping

CpG methylation: Blocked by some combinations of overlapping

 

Percent Activity in MINOTECH Buffers
 L M  H  SH  A  K 
 75-100 100  25-50  10-25  75-100  100 
 
 
General reaction mixture:
10U Sfi I 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