Recognition Sequence :
GTATCC(N)6↑
CATAGG(N)5↓
Unit Definition :
One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 37°C in a total reaction volume of 50 μl.
Reaction Temperature :
37°C
Storage Buffer :
10 mM Tris-HCl (pH 7.5); 100 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA, 0.05% Triton X-100, 50% glycerol
Ligation :
After 5-fold overdigestion with enzyme ~10% of DNA fragments can be ligated with T4 DNA Ligase and recut.
Source :
Bacillus sphaericus
Working buffer :
Y (33 mM Tris-acetate (pH 7.9 at 25°C); 10 mM magnesium acetate; 66 mM potassium acetate; 1 mM DTT.)
Non-specific hydrolisis :
No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 4 u.a. of enzyme for 16 hours at 37°C.
Concentration, u.a./ml :
2000-5000
Inactivation :
20min Under 65°C
Reagents Supplied :
10 X SE-buffer Y
Notes :
When using a buffer other than the optimal (suppied) SE Buffer, it may be necessary to add more enzyme to achieve complete digestion.
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