Fok I

Description

Recognition site and hydrolysis position:

Source: An E.coli strain that carries the cloned Fok I gene from Flavobacterium okeanokoites
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.
Assayed on:
Lambda DNA
Optimal SE-buffer: SE-buffer Y
Enzyme activity (%):

Storage conditions: 10 mM Tris-HCl (pH 7.5); 50 mM NaCl; 0,1 mM EDTA; 1 mM DTT; 200 μg/ml BSA; 50% glycerol; Store at -20°C.
Ligations:
After 2-fold overdigestion with enzyme more than 95% of the DNA fragments can be ligated and recut.
Non-specific hydrolisis:
No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 1 u.a. of enzyme for 16 hours at 37°C Reagents Supplied with Enzyme 10 X SE-buffer Y
Reagents Supplied with Enzyme:
Methylation sensitivity: not tested
Notes: Overdigestions of > 5 units of Fok I per 1μg of DNA and incubations > 2 hours are not recommended.
References:
Sugisaki, H., Kanazawa, S., Gene 16: 73-78 (1981).