BslF I

Description

Recognition site and hydrolysis position:

Source: Bacillus stearothermophilus Fl
Unit definition:
One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 37°C in a total reaction volume of 50 μl.
Assayed on:
Lambda DNA
Optimal SE-buffer: SE-buffer Y + BSA
Enzyme activity (%):

Storage conditions: 10 mM Tris-HCl (pH 7.5); 250 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 50% glycerol; Store at -20°C.
Ligations:
After 3-fold overdigestion with enzyme 90% of DNA fragments can be ligated and 95% of these can be recut.
Non-specific hydrolisis:
No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 2 u.a. of enzyme for 16 hours at 37°C. Reagents Supplied with Enzyme 10 X SE-buffer Y, BSA
Reagents Supplied with Enzyme:
Methylation sensitivity: not tested
Notes: High enzyme concentration may result in star activity.
Long incubation with BSA is not recommend.
To obtain 100% activity, BSA should be added to the 1x reaction mix to a final concentration of 100 μg/ml.
There is DNA-methyltransferase activity in presence of SAM. It is maximum at 48°C. In presence of 10mM MgCl₂ enzyme both modifies and hydrolyzes DNA. If MgCl₂ is absent enzyme modifies DNA only. And that DNA become proof against BslFI.
BslF I also cleaves the sequence GGGAC(11/15).
References:
Nadeev, A.N., Kashirina, J.G., Nayakshina, T.N., Dedkov, V.S., Mezentseva, N.V., Tomilova, J.E., Degtyarev, S.K.; Unpublished observations. 2004