Source: Bacillus stearothermophilus BA
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 65°C in a total reaction volume of 50 μl.
Optimal SE-buffer: W (10 mM Tris-HCl (pH 8.5 at 25°C); 10 mM MgCl2; 100 mM NaCl; 1 mM DTT.) + BSA
Activity in SEBuffers:
SEBuffer B 25 - 50%
SEBuffer G 25 - 50%
SEBuffer O 75 -100%
SEBuffer W 100%
SEBuffer Y 25 - 50%
SEBuffer ROSE 50%
When using a buffer other than the optimal (suppied) SEBuffer, it may be necessary to add more enzyme to achieve complete digestion.
Optimal temperature: 65°C
Storage conditions: 10 mM Tris-HCl (pH 7.5); 50 mM KCl; 0.1 mM EDTA; 10 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol.
Store at: -20°C.
Ligation: After 10-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 65°C.
Reagents supplied with enzyme: 10 X SE-buffer W, BSA
Methylation sensitivity: Blocked by CG methylation.
Inactivation: 20 minutes under 80°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.
BstBA I - YAC↑GTR - RTG↓CAY - Restriction Endnuclease from Sibenzyme
* availibility of sample size may be limited
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