Fae I

Description

Recognition site and hydrolysis position:

Source: Flavobacterium aquatile N3
Unit definition:
One unit of the enzyme is the amount required to hydrolyze 1 μg of pUC19 DNA in 1 hour at 37°C in a total reaction volume of 50 μl.
Assayed on:
pUC19 DNA
Optimal SE-buffer: SE-buffer FaeI
Enzyme activity (%):

Storage conditions: 10 mM Tris-HCl (pH 7.5); 50 mM KCl; 0,1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol; Store at -20°C.
Ligations:
After 3-fold overdigestion with enzyme more then 90% of the DNA fragments can be ligated with T4 DNA Ligase at 16°C and recut.
Non-specific hydrolisis:
No nonspecific activity was detected after incubation of 1 μg of pUC19 DNA with 2 u.a. of enzyme for 16 hours at 37°C. Reagents Supplied with Enzyme 10 X SE-buffer FaeI, BSA
Reagents Supplied with Enzyme:
Methylation sensitivity: Blocked by C^mATG methylation
Notes: To obtain 100% activity, BSA should be added the 1x reaction mix to a final concentration of 100 μg/ml.
Do not use BSA for long incubation.
References:
Chernukhin, V.A., Kileva, E.V., Tomilova, J.E., Dedkov, V.S.. Unpublished observations (2006).