SgrBI (SacII isoschizomer)
* Size:


(SacII isoschizomer)

Technical Brochure Technical Brochure
SgrBI is a restriction enzyme purified from Streptomyces griseus.


Unit substrate: Lambda DNA (HindIII digest).


Unit calculation assay conditions: 10 mM Tris-HCl (pH 7.9@ 25°C), 10 mM MgCl2, 1 mM dithiothreitol, 0.1% Triton X-100, 100 μg/ml BSA. Incubate at 37°C.


Absence of contaminants: 400 units of SgrB I do not produce any unspecific cleavage products after 16 hrs incubation with 1 μg of λ DNA (HindIII digest) at 37°C. After 100-fold overdigestion with SgrBI, greater than 98% of the DNA fragments can be ligated and recut with this enzyme.


Storage buffer: 50 mM NaCl, 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: 65oC for 20 minutes.


Methylation Sensitivity:

dam methylation: Not sensitive

dcm methylation: Not sensitive

CpG methylation: Blocked


Note: Particular sites in λ and φΧ174 DNAs are difficult to cleave with SgrB I, as well as with its prototype Sac II.

Reference: Rina, M., Pagomenou, M. and V, Bouriotis (1991) Nucleic Acids Res. 19, 6342.


Percent Activity in MINOTECH Buffers
 L SH 
 75-100 75  50-75  25-50  <10  100 
General reaction mixture:
10U SgrBI 1μl
10x USgrBI or K buffer*  2μl 
DNA substrate  <1μg
Sterile ultrapure water  Up to 20 μl
Incubate for 15 min at 37oC  
*In the case of USgrBI buffer we recommend the addition of BSA to a final concentration of 100 μg/ml. 


Reagents supplied: 10x USgrBI and 10x K buffer