|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 (%)|
|75 - 100||100||25 - 50||25 - 50||75 - 100||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.|
|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).|
|Theoretical diagrams of DNA digestion by this enzyme for the most known DNA substrates:|
To view the fragments length values please point mouse cursor over diagram
M - ladder, 1 - Adeno-2 DNA, 2 - Lambda DNA, 3 - T7 DNA, 4 - pUC19, 5 - pBR322