BstBA I

Description

Recognition site and hydrolysis position:

Source: Bacillus stearothermophilus BA
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.
Assayed on:
Lambda DNA
Optimal SE-buffer: SE-buffer W + BSA
Enzyme activity (%):

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.
Ligations:
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
Reagents Supplied with Enzyme:
Methylation sensitivity: Blocked by CG methylation.
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.
References:
Abdurashitov, M.A., Shinkarenko, N.M., Dedkov, V.S., Shevchenko, A.V., Degtyarev, S.K. Unpublished observations (1996).