Recognition Sequence :
GKGCM↑C
C↓MCGKG
Unit Definition :
One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 55°C in a total reaction volume of 50 μl.
Reaction Temperature :
55°C
Storage Buffer :
10 mM Tris-HCl (pH 7.5); 50 mM KCl; 0,1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; and 50% glycerol
Ligation :
After 5-fold overdigestion with enzyme 80% of the DNA fragments can be ligated. Of these, 95% can be recut.
Source :
Bacillus stearothermophilus S
Working buffer :
G (10 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 50 mM NaCl; 1 mM DTT.) + BSA
Non-specific hydrolisis :
No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 10 u.a. of enzyme for 16 hours at 55°C.
Concentration, u.a./ml :
10000
Inactivation :
20min Under 65°C
Methylation sensitivity :
Not blocked by overlapping dcm-methylation (CmCWGG): GKGCCCWGG
Blocked by GKG(5mC)MC methylation
Reagents Supplied :
10 X SE-buffer G, BSA
Notes :
To obtain 100% activity, BSA should be added to the 1 x reaction mix to a final concentration of 100 μg/ml.
Do not use BSA for long incubation.
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