FauND I

Enzyme nameFauND I
PrototypeNdeI
Known analogsNdeI
SKUSE-E009
Turbo versionAvailable
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'… CA▼TATG …3'
3'… GTAT▲AC …5'
SourceAn E.coli strain that carries the cloned FauND I gene from Flavobacterium aquatili ND
Optimal bufferSE-buffer Y (33 mM Tris-acetate (pH 7.9 at 25°C); 10 mM magnesium acetate; 66 mM potassium acetate; 1 mM DTT.)
Optimal temperature37 °C
Inactivation temperature65 °C
Buffer activity
BGOWYROSE
50751050100100
Unit definitionOne 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 onLambda DNA
Storage conditions10 mM Tris-HCl (pH 7.5); 50 mM KCl; 0,1 mM EDTA; 1mM DTT; 200 μg/ml BSA, 50% glycerol; Store at -20°C.
LigationAfter 10-fold overdigestion with enzyme 80% of the DNA fragments can be ligated and recut. In the presence of 10% PEG
Nonspecific hydrolysisNo nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 10 u.a. of enzyme for 16 hours at 37°C.
Methylation sensitivitynot tested
Reagents supplied with enzyme10 X SE-buffer Y, BSA
NotesSensitive to impurities present in some DNA preparations. For example, DNA purified by standard miniprep procedures is cleaved at lower rates. To obtain 100% activity, BSA should be added to the 1 x reaction mix to a final concentration of 100 μg/ml. Do not use BSA for long incubation.
References
  2. Unpublished data Shevchenko, A.V., Belichenko, O.A., Myakisheva, T.V., Dedkov, V.S., Abdurashitov, M.A., Degtyarev, S.K Unpublished observations (1995).
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