Kpn I

Enzyme nameKpn I
PrototypeKpnI
Known analogsAcc65 I, Asp718I
SKUSE-E079
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'… GGTAC▼C …3'
3'… C▲CATGG …5'
SourceAn E.coli strain that carries the cloned Kpn I gene from Klebsiella pneumonia
Optimal bufferSE-buffer B (10 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 1 mM DTT.)
Optimal temperature37 °C
Inactivation temperature80 °C
Buffer activity
BGOWYROSE
1002525257550
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); 50 mM NaCl; 0,1 mM EDTA; 1 mM DTT; 200 μg/ml BSA; 50% glycerol; Store at -20°C.
LigationAfter 20-fold overdigestion with enzyme more than 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 blocked by overlapping Dcm methylation (C m CWGG) GGTA CC WGG
Reagents supplied with enzyme10 X SE-buffer B, BSA
NotesHigh enzyme concentration may result in star activity. Long incubation with BSA is not recommended due to star activity. To obtain 100% activity, BSA should be added to the 1 x reaction mix to a final concentration of 100 μg/ml.
References
  2. External publication Smith, D.I., Blattner, F.R., Davies, J. Nucleic Acids Res. 3: 343-353 (1976).
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