Source: Bacillus stearothermophilus Fl
Assayed on: Lambda DNA
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.
Optimal SE-buffer Y (33 mM Tris-acetate (pH 7.9 at 25°C); 10 mM magnesium acetate; 66 mM potassium acetate; 1 mM DTT.) + BSA
Activity in SEBuffers:
SEBuffer B 25 - 50%
SEBuffer G 25 - 50%
SEBuffer O 10 - 25%
SEBuffer W 25 - 50%
SEBuffer Y 100%
SEBuffer ROSE 50%
When using a buffer other than the optimal (suppied) SEBuffer, it may be necessary to add more enzyme to achieve complete digestion.
Optimal temperature: 37°C
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.
Ligation: 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
Methylation sensitivity: not tested
Inactivation: 20 minutes under 80°C
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).
BslF I - GGGAC(N)10↑ - CCCTG(N)14↓ - Restriction Endnuclease from Sibenzyme
* availibility of sample size may be limited
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