Recognition Sequence :
GGGAC(N)10↑
CCCTG(N)14↓
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.
Reaction Temperature :
37°C
Storage Buffer :
10 mM Tris-HCl (pH 7.5); 250 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 50% glycerol
Ligation :
After 3-fold overdigestion with enzyme 90% of DNA fragments can be ligated and 95% of these can be recut.
Source :
Bacillus stearothermophilus Fl
Working buffer :
Y (33 mM Tris-acetate (pH 7.9 at 25°C); 10 mM magnesium acetate; 66 mM potassium acetate; 1 mM DTT.) + BSA
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.
Concentration, u.a./ml :
1000
Inactivation :
20min Under 80°C
Reagents Supplied :
10 X SE-buffer Y, BSA
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).
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