Pct I

Enzyme namePct I
PrototypeBsmI
Known analogsBsaMI, BsmI, Mva1269I
SKUSE-E045
Turbo versionNot available
High-concentration versionNot available
Restriction typeType IISiType IIS enzymes recognize asymmetric DNA sequences and cleave outside the recognition site at a defined distance. They are widely used in Golden Gate assembly.
Recognition site
5'… GAATGCN▼ …3'
3'… CTTAC▲GN …5'
SourcePlanococcus citreus SM
Optimal bufferSE-buffer O (50 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 100 mM NaCl; 1 mM DTT.)
Optimal temperature37 °C
Inactivation temperature65 °C
Buffer activity
BGOWYROSE
2550100751050
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); 250 mM NaCl; 0,1 mM EDTA; 7 mM 2-mercaptoethanol; 100 μg/ml BSA; and 50% glycerol. Store at -20°C.
LigationAfter 10-fold overdigestion with enzyme >90% 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 37°C
Methylation sensitivitynot tested
Reagents supplied with enzyme10 X SE-buffer O
References
  2. Unpublished data Dedkov, V.S., Nayakshina, T.N., Popichenko, D.V., Degtyarev, S.K Unpublished observations (2000).
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