Source: Bacillus species AC
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: O (50 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 100 mM NaCl; 1 mM DTT.) + BSA
Activity in SEBuffers:
SEBuffer B 10 - 25%
SEBuffer G 25 - 50%
SEBuffer O 100%
SEBuffer W 75 - 100%
SEBuffer Y 10 - 25%
SEBuffer ROSE 100%
When using a buffer other than the optimal (suppied) SEBuffer, it may be necessary to add more enzyme to achieve complete digestion.
Optimal temperature: 37°C
Storage conditions: 10 mM KH2PO4(pH 7.2); 100 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol.
Store: at -20°C.
Ligation: After 5-fold overdigestion with enzyme more than 95% of the Lambda DNA fragments can be ligated with T4 DNA Ligase at 16°C and 50% of these can be recut.
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 O, BSA
Methylation sensitivity: Blocked by CG methylation.
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.
BspACI has a non-palindromic recognition site.
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