Recognition site | GTY↑RAC CAR↓YTG |
Source | An E.coli strain, that carries the cloned gene HindII from Haemophilus influenzae |
Assayed on | Lambda DNA |
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. |
Optimal SE-buffer | G (10 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 50 mM NaCl; 1 mM DTT.) + BSA |
Enzyme activity (%) | B | G | O | W | Y | Rose | 75 - 100 | 100 | 25 - 50 | 25 - 50 | 75 - 100 | 50 |
|
Optimal temperature | 37oC |
Storage conditions | 10 mM Tris-HCl (pH 7.5); 200 mM NaCl; 0.1 mM EDTA; 1 mM DTT; 200 μg/ml BSA; 50% glycerol. Store at -20°C. |
Ligation | After 10-fold overdigestion with enzyme 60% of the DNA fragments can be ligated and recut. In the presence of 10%PEG ligation is better. |
Non-specific hydrolisis | No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 20 u.a. of enzyme for 16 hours at 37°C.
|
Reagents Supplied with Enzyme |
10 X SE-buffer G, BSA.
|
Methylation sensitivity | not tested |
Inactivation 20 minutes under | 65oC |
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.
|
Quality control | Restriction Endonucleases: Quality Control |
MSDS: | Download MSDS as PDF |
References: | Kelly, T.J. Jr., Smith, H.O.
J. Mol. Biol. 51: 393-409 (1970).
|