Recognition Sequence :
CA↑TATG
GTAT↓AC
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); 50 mM KCl; 0,1 mM EDTA; 1mM DTT; 200 μg/ml BSA, 50% glycerol
Ligation :
After 10-fold overdigestion with enzyme 80% of the DNA fragments can be ligated and recut. In the presence of 10% PEG ligation is better.
Source :
An E.coli strain that carries the cloned FauND I gene from Flavobacterium aquatili ND
Working buffer :
Y (33 mM Tris-acetate (pH 7.9 at 25°C); 10 mM magnesium acetate; 66 mM potassium acetate; 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 37°C.
Concentration, u.a./ml :
10000
Inactivation :
20min Under 65°C
Reagents Supplied :
10 X SE-buffer Y, BSA
Notes :
Sensitive to impurities present in some DNA preparations. For example, DNA purified by standard miniprep procedures is cleaved at lower rates.
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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