BstC8 I

Enzyme nameBstC8 I
PrototypeCac8I
Known analogsCac8I
SKUSE-E305
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'… GCN▼NGC …3'
3'… CGN▲NCG …5'
SourceBacillus stearothermophilus C8
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 temperature55 °C
Inactivation temperature80 °C
Buffer activity
BGOWYROSE
10255075100100
Unit definitionOne unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 55°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; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol; Store at -20°C.
LigationAfter 10-fold overdigestion with enzyme more than 95% of the DNA fragments can be ligated and recut.
Nonspecific hydrolysisNo nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 20 u.a. of enzyme for 16 hours at 55°C.
Methylation sensitivityBlocked by GC methylation: 5`-G(5mC)NNGC-3`/3`-CGNN(5mC)G-5`.
Reagents supplied with enzyme10 X SE-buffer Y
NotesAt 37°C activity is 50% from maximum.
References
  2. Unpublished data Abdurashitov, M.A., Dedkov, V.S., Mikhnenkova, N.A., Kileva, E.V., Popichenko, D.V., Degtyarev, S.K Unpublished observations (2001).
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