Kpn I

Description

Recognition site and hydrolysis position:

Source: An E.coli strain that carries the cloned Kpn I gene from Klebsiella pneumonia
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
Optimal SE-buffer: SE-buffer B
Enzyme activity (%):

Storage conditions: 10 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.
Ligations:
After 20-fold overdigestion with enzyme more than 90% of the DNA fragments can be ligated and recut.
Non-specific hydrolisis:
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. Reagents Supplied with Enzyme 10 X SE-buffer B, BSA
Reagents Supplied with Enzyme:
Methylation sensitivity: Not blocked by overlapping Dcm methylation (C^mCWGG): GGTACCWGG
Notes: High 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:
Smith, D.I., Blattner, F.R., Davies, J. Nucleic Acids Res. 3: 343-353 (1976).
Abdurashitov M.A., Tomilov V.N., Chernukhin V.A., Gonchar D. A., Degtyarev S. Kh. Comparative analysis of human chromosomal DNA digestion with restriction endonucleases in vitro and in silico. // Translated from “Medical genetics” V.6, No 8, pp 29-36, 2007