Pct I

Enzyme name Pct I
Prototype BsmI
Known analogs BsaMI, BsmI, Mva1269I
SKU SE-E045
Turbo version Not available
High-concentration version Not available
Restriction type Type 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'
Source Planococcus citreus SM
Optimal buffer SE-buffer O (50 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 100 mM NaCl; 1 mM DTT.)
Optimal temperature 37 °C
Inactivation temperature 65 °C
Buffer activity
BGOWYROSE
2550100751050
Unit definition One 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 on Lambda DNA
Storage conditions 10 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.
Ligation After 10-fold overdigestion with enzyme >90% of the DNA fragments can be ligated and recut
Nonspecific hydrolysis No 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 sensitivity not tested
Reagents supplied with enzyme 10 X SE-buffer O
Notes
References
  2. Unpublished data Dedkov, V.S., Nayakshina, T.N., Popichenko, D.V., Degtyarev, S.K Unpublished observations (2000).
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