Description
Recognition site and hydrolysis position:
| GTYRAC | GTY↑RAC |
| CARYTG | CAR↓YTG |
Source: An E.coli strain, that carries the cloned gene HindII from Haemophilus influenzae
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.
Assayed on:
Lambda DNA
Optimal SE-buffer: SE-buffer G
Enzyme activity (%):
| B | G | O | W | Y | ROSE |
| 75 | 100 | 25 | 25 | 75 | 50 |
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.
Ligations:
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 10 u.a. of enzyme for 16 hours at 37°C.Reagents Supplied with Enzyme
10 X SE-buffer G, BSA.
Reagents Supplied with Enzyme:
Methylation sensitivity: not tested
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.
References:
Kelly, T.J. Jr., Smith, H.O. J. Mol. Biol. 51: 393-409 (1970).
