BstAF I

Enzyme name BstAF I
Prototype AflII
Known analogs AflII, BfrI, BspTI, Bst98I, MspCI, Vha464I
SKU SE-E135
Turbo version Not available
High-concentration version Not available
Restriction type Type IIPiType IIP enzymes are classical Type II restriction enzymes with palindromic recognition sequences and symmetric cleavage sites.
Recognition site
5'… C▼TTAAG …3'
3'… GAATT▲C …5'
Source Bacillus stearothermophilus AF
Optimal buffer SE-buffer W (10 mM Tris-HCl (pH 8.5 at 25°C); 10 mM MgCl2; 100 mM NaCl; 1 mM DTT.)
Optimal temperature 55 °C
Inactivation temperature 80 °C
Buffer activity
BGOWYROSE
10257510025100
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
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.
Ligation 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
Nonspecific hydrolysis 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.
Methylation sensitivity not tested
Reagents supplied with enzyme 10 X SE-buffer W
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

No publications found.
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