| Recognition site | GGGAC(N)10↑
CCCTG(N)14↓ |
| Source | Bacillus stearothermophilus Fl |
| Assayed on | Lambda DNA |
| Unit | 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 |
| Enzyme activity (%) | | B | G | O | W | Y | | 25 - 50 | 25 - 50 | 10 - 25 | 25 - 50 | 100 |
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| Optimal temperature | 37oC |
| 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% 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.
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| Reagents Supplied with Enzyme |
10 X SE-buffer Y, BSA |
| Methylation sensitivity | not tested |
| Inactivation 20 minutes under | 80oC |
| 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). |
| MSDS: | Download MSDS as PDF |
| 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Unpublished observations. 2004
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Theoretical diagrams of DNA digestion by this enzyme for the most known DNA substrates:
To view the fragments length values please point mouse cursor over diagram |
M - ladder, 1 - Adeno-2 DNA, 2 - Lambda DNA, 3 - T7 DNA, 4 - pUC19, 5 - pBR322
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