BstAF I

Description

Recognition site and hydrolysis position:

Source: Bacillus stearothermophilus AF
Unit definition:
One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 55°C in a total reaction volume of 50 μl.
Assayed on:
Lambda DNA
Optimal SE-buffer: SE-buffer W
Enzyme activity (%):

Storage conditions: 10 mM Tris-HCl (pH 7.5); 200 mM KCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol. Store at -20°C.
Ligations:
After 5-fold overdigestion with enzyme ~40% of the DNA fragments can be ligated and 95% of these can be recut. In the presence of 10% PEG ligation is better.
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 55°C. Reagents Supplied with Enzyme 10 X SE-buffer W
Reagents Supplied with Enzyme:
Methylation sensitivity: not tested
Notes: Do not use BSA for long incubation.
To obtain 100% activity, BSA should be added to the 1 x reaction mix to a final concentration of 100 μg/ml.
References: