Pst I

Enzyme namePst I
PrototypePstI
Known analogsBspMAI
SKUSE-E109
Turbo versionAvailable
High-concentration versionAvailable
Restriction typeType IIPiType IIP enzymes are classical Type II restriction enzymes with palindromic recognition sequences and symmetric cleavage sites.
Recognition site
5'… CTGCA▼G …3'
3'… G▲ACGTC …5'
SourceAn E.coli strain that carries the cloned Pst I gene from Providencia stuartii
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 temperature80 °C
Buffer activity
BGOWYROSE
1025100252550
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); 200 mM NaCl; 0,1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol. Store at -20°C.
LigationAfter 20-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 40 u.a. of enzyme for 16 hours at 37°C.
Methylation sensitivitynot tested
Reagents supplied with enzyme10 X SE-buffer O, BSA (except E109T and E110T).
NotesHigh enzyme concentration may result in star activity. 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
  1. Y.N. Boikov, V.E. Repin, S.Kh. Degtyarev, V.A. Viesture, O.A. Stengrevitzs Providencia stuartii - a strain producer of restriction endonuclease PstI. // Soviet Union patent SU 1472493 (1989). (In Russian)
  3. External publication Smith, D.I., Blattner, F.R., Davies, J. Nucleic Acids Res. 3: 343-353 (1976).
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