Fae I

Enzyme nameFae I
PrototypeNlaIII
Known analogsHin1II, Hsp92II, NlaIII, CviAII, Fat I
SKUSE-E495
Turbo versionNot available
High-concentration versionNot available
Restriction typeType IIPiType IIP enzymes are classical Type II restriction enzymes with palindromic recognition sequences and symmetric cleavage sites.
Recognition site
5'… CATG▼ …3'
3'… ▲GTAC …5'
SourceFlavobacterium aquatile N3
Optimal bufferSE-buffer FaeI (33 mM Tris-acetate (pH 8.3 at 25°C); 10 mM magnesium acetate; 66 mM potassium acetate; 1 mM DTT.)
Optimal temperature37 °C
Inactivation temperature65 °C
Buffer activity
BGOWYROSE
2550101075100
Unit definitionOne 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 onpUC19 DNA
Storage conditions10 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.
LigationAfter 3-fold overdigestion with enzyme more then 90% of the DNA fragments can be ligated with T4 DNA Ligase at 16°C and recut.
Nonspecific hydrolysisNo nonspecific activity was detected after incubation of 1 μg of pUC19 DNA with 2 u.a. of enzyme for 16 hours at 37°C.
Methylation sensitivityBlocked by C m ATG methylation
Reagents supplied with enzyme10 X SE-buffer FaeI, BSA
NotesTo 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
  2. Unpublished data Chernukhin, V.A., Kileva, E.V., Tomilova, J.E., Dedkov, V.S Unpublished observations (2006)
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