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SmaI
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Sma I Technical Brochure Technical Brochure
SmaI is a restriction enzyme purified from Serratia marcescens (ATCC 49779).

 

Unit substrate: Lambda DNA (HindIII digest).

 

Unit calculation assay conditions: 50 mM potassium acetate, 20 mM Tris-acetate (pH 7.9 @ 25°C), 10 mM magnesium acetate, 1 mM dithiothreitol, 100 μg/ml BSA. Incubate at 25oC.

 

Absence of contaminants: 150 units of SmaI do not produce any unspecific cleavage products after 16 hrs incubation with 1 μg of λ DNA (HindIII digest) at 25oC. After 50-fold overdigestion with SmaI, 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 7.4), 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

 

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

 

Reagents supplied: 10x A and 10x K buffer