Activity in SEBuffers:

SEBuffer B       10 - 20%

SEBuffer G             100%

SEBuffer O       25 - 50%

SEBuffer W       10 - 25%

SEBuffer Y        50 - 75%

SEBuffer ROSE      100%

When using a buffer other than the optimal (suppied) SEBuffer, it may be necessary to add more enzyme to achieve complete digestion. 

Optimal temperature: 55°C

Storage conditions: 10 mM Tris-HCl (pH 7.5); 50 mM KCl; 0,1 mM EDTA; 1 mM DTT; 200 μg/ml BSA; and 50% glycerol.

Store: at -20°C.

Ligation: After 2-fold overdigestion with enzyme more than 90% of the DNA fragments can be ligated and recut.

Non-specific hydrolisis: No nonspecific activity was detected after incubation of 1 μg of DNA with 3 u.a. of enzyme for 16 hours at 55°C.

Reagents supplied with Enzyme: 10 X SE-buffer G

Methylation sensitivity: Blocked by mCATG methylation

Inactivation: 20 minutes under 65°C

Notes: The minimum number of units that resulted in complete digestion of 1 μg of substrate DNA in 16 hours is 0,25. FatI cleaves linear plasmid DNA at a rate 1.5-2 times higher than supercoiled plasmid DNA.