Recognition site and hydrolysis position:
Source: Actinobacillus suis NH
One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA (HindIII-digest) in 1 hour at 37°C in a total reaction volume of 50 μl.
Lambda DNA (HindIII-digest)
Optimal SE-buffer: SE-buffer Y + BSA
Enzyme activity (%):
Storage conditions: 10 mM Tris-HCl (pH 7.5); 250 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 100 μg/ml BSA; 50% glycerol. Store at -20°C
After 20-fold overdigestion with enzyme more than 90% of the DNA fragments can be ligated and recut
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:
Methylation sensitivity: not tested
Notes: To obtain 100% activity, BSA should be added to the 1x reaction mix to a final concentration of 100 μg/ml.
Do not use BSA for long incubation.
Dedkov, V.S., Degtyarev, S.Kh. Biol. Chem. 379: 573-574 (1998).