Recognition Sequence :
C↑TTAAG
GAATT↓C
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); 200 mM KCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol.
Ligation :
After 5-fold overdigestion with enzyme ~40% of the DNA fragments can be ligated and 95% of these can be recut. In the presence of 10% PEG ligation is better.
Source :
Bacillus stearothermophilus AF
Working buffer :
W (10 mM Tris-HCl (pH 8.5 at 25°C); 10 mM MgCl2; 100 mM NaCl; 1 mM DTT.) + BSA
Non-specific hydrolisis :
No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 40 u.a. of enzyme for 16 hours at 55°C.
Concentration, u.a./ml :
20000
Inactivation :
20min Under 80°C
Reagents Supplied :
10 X SE-buffer W
Notes :
Do not use BSA for long incubation.
To obtain 100% activity, BSA should be added to the 1 x reaction mix to a final concentration of 100 μg/ml.
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