BslF I - GGGAC(N)10↑ - CCCTG(N)14↓

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).