Recognition Sequence :
CTGAAG(N)16↑
GACTTC(N)14↓
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; 1 mM DTT; 200 μg/ml BSA; 50% glycerol
Ligation :
After 2-fold overdigestion with enzyme about 80% of the DNA fragments can be ligated. Of these, 80% can be recut.
Source :
An. E.coli strain that carries the cloned Acu I gene from Acinetobacter calcoaceticus SRW4
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 + SAM
Non-specific hydrolisis :
No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 1 u.a. of enzyme for 16 hours at 37°C.
Concentration, u.a./ml :
1000
Inactivation :
20min Under 65°C
Reagents Supplied :
10 X SE-buffer Y, BSA, SAM
Notes :
High enzyme concentration may result in star activity.
To obtain 100% activity, BSA should be added to the 1x reaction mix to a final concentration of 100 μg/ml and SAM should be added to a final concentration 10 μM. It is possible to use a final SAM concentration of 10-64 μM. In this case 32 mM SAM solution should be diluted in 3200-500 times, correspondingly. For the small volume of reaction mix (10-20 μl) the stock SAM solution may be previously diluted up to 1 mM (in 32 times) with 5 mM solution of H2SO4 or sterile water and stored at -20°C.
Do not use BSA for long incubation.
Our Products Cannot Be Used As Medicines Directly For Personal Use.
PDF