Source: Bacillus species 19
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: 2W (20 mM Tris-HCl (pH 8,5 at 25°C); 10 mM MgCl2; 200 mM NaCl; 1 mM DTT) + BSA
Activity in SEBuffers:
SEBuffer B 0 - 10%
SEBuffer G 10 - 25%
SEBuffer O 50 - 75%
SEBuffer W 75 -100%
SEBuffer Y 10 - 25%
SEBuffer ROSE 5%
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 Tris-HCl (pH 7.5); 50 mM KCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 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 40 u.a. of enzyme for 16 hours at 37°C.
Reagents Supplied with Enzyme: 10 X SE-buffer 2W, BSA
Methylation sensitivity: Bsp19I cuts hemimethylated site
and doesn't cut methylated sites
Inactivation: 20 minutes under 65oC
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.
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