Source: Actinobacillus suis NH
Assayed on: Lambda DNA (HindIII-digest)
Unit definition: 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.
Optimal SE-buffer Y (33 mM Tris-acetate (pH 7.9 at 25°C); 10 mM magnesium acetate; 66 mM potassium acetate; 1 mM DTT.) + BSA
Activity in SEBuffers:
SEBuffer B 75 -100%
SEBuffer G 50 - 75%
SEBuffer O 0 - 10%
SEBuffer W 0 - 10%
SEBuffer Y 100%
SEBuffer ROSE 25%
When using a buffer other than the optimal (supplied) SEBuffer, it may be necessary to add more enzyme to achieve complete digestion.
Optimal temperature: 37°C
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
Ligation: After 20-fold overdigestion with enzyme more than 90% of the DNA fragments can be ligated and recut
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 Y, BSA
Methylation sensitivity: not tested
Inactivation: 20 minutes under 65°C
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.
AsuNH I - G↑CTAGC - CGATC↓G
Restriction Endnuclease from Sibenzyme
* availibility of sample size may be limited
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