Btr I

Description

Recognition site and hydrolysis position:

Source: Bacillus stearothermophilus SE-U62
Unit definition:
One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 60°C in a total reaction volume of 50 μl.
Assayed on:
Lambda DNA
Optimal SE-buffer: SE-buffer O
Enzyme activity (%):

Storage conditions: 10 mM Tris-HCl (pH 7.5); 50 mM KCl; 0,1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; and 50% glycerol; Store at -20°C.
Ligations:
After 3-fold overdigestion with enzyme 80% of DNA fragments can be ligated. Of these 90% can be recut.
Non-specific hydrolisis:
No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 5 u.a. of enzyme for 16 hours at 60°C. Reagents Supplied with Enzyme 10 X SE-buffer O, BSA
Reagents Supplied with Enzyme:
Methylation sensitivity: not tested
Notes: High enzyme concentration may result in star activity.
To obtain 100% activity, BSA should be added to the 1 x reaction mix to a final concentration of 100 μg/ml.
Do not use BSA for long incubation.
References:
Degtyarev, S.Kh., Belichenko, O.A., Lebedeva, N.A., Dedkov, V.S., Abdurashitov, M.A. Nucleic Acids Res. 28: p56 (2000).