Pci I

Enzyme namePci I
PrototypeBspLU11I
Known analogsBspLU11I, PscI
SKUSE-E275
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'… A▼CATGT …3'
3'… TGTAC▲A …5'
SourceAn E.coli strain that carries the cloned Pci I gene from Planococcus citreus SE-F45
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
5075100755050
Unit definitionOne unit of the enzyme is the amount required to hydrolyze 1 μg of T7 DNA in 1 hour at 37°C in a total reaction volume of 50 μl.
Assayed onT7 DNA
Storage conditions10 mM Tris-HCl (pH 7.5); 250 mM NaCl; 0,1 mM EDTA; 7 mM 2-mercaptoethanol; 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 T7 DNA with 10 u.a. of enzyme for 16 hours at 37°C.
Methylation sensitivityNot blocked by AC m ATGT methylation. Blocked by m ACATGT methylation.
Reagents supplied with enzyme10 X SE-buffer O
NotesThe minimum number of units that resulted in complete digestion of 1 μg of substrate DNA in 16 hours is 0,13.
References
  2. Unpublished data Abdurashitov, M.A., Nayakshina, T.N., Lebedeva, N.A., Dedkov, V.S., Degtyarev, S.K Unpublished observations (1998).
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