Fae I

Enzyme name Fae I
Prototype NlaIII
Known analogs Hin1II, Hsp92II, NlaIII, CviAII, Fat I
SKU SE-E495
Turbo version Not available
High-concentration version Not available
Restriction type Type 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'
Source Flavobacterium aquatile N3
Optimal buffer SE-buffer FaeI (33 mM Tris-acetate (pH 8.3 at 25°C); 10 mM magnesium acetate; 66 mM potassium acetate; 1 mM DTT.)
Optimal temperature 37 °C
Inactivation temperature 65 °C
Buffer activity
BGOWYROSE
2550101075100
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
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.
Ligation 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.
Nonspecific hydrolysis 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.
Methylation sensitivity Blocked by C m ATG methylation
Reagents supplied with enzyme 10 X SE-buffer FaeI, BSA
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
  2. Unpublished data Chernukhin, V.A., Kileva, E.V., Tomilova, J.E., Dedkov, V.S Unpublished observations (2006)
// Код для некролога