Kpn I from Klebsiella pneumoniae buffered aqueous glycerol solution

Properties

Related Categories	3.1.x.x Acting on esters, 3.x.x.x Hydrolases, Application Index, Biochemicals and Reagents, Enzyme Class Index, Enzymes, Inhibitors, and Substrates, Molecular Biology, Molecular Biology Enzymes, Molecular Biology Tested, Restriction Endonucleases, Restriction Enzymes More...
form	buffered aqueous glycerol solution
concentration	10,000 units/mL
shipped in	wet ice
storage temp.	−20°C

Description

Other Notes

Supplied with 10x Restriction Endonuclease Buffer SL (B3782).

Comment: Kpn I shows abnormal DNA cleaving patterns (star activity) in reactions containing high (>5%) glycerol, >5% (v/v); excess enzyme and high pH (>8.0).
The enzyme will not cleave DNA containing 3′-residue of 5-methylcytosine.

Specificity

Recognition sequence: 5′-GGTAC/C-3′
Ligation and recutting results: After 2-10-fold Kpn I overdigestion of 1 μg λ DNA substrate, results in 100% cutting, >95% of fragments can be ligated, and >95% recut.
Heat inactivation: 65 °C for 15 minutes.

Physical form

Solution in 20 mM Tris-HCl, pH 7.5, 0.1 mM EDTA, 50 mM NaCl, 10 mM 2-mercaptoethanol, 50% glycerol (v/v), 0.01% polydocanol (v/v) at 4°C

Application

KpnI is a DNA restriction enzyme that is used in molecular biology research to cleave DNA at the recognition sequence 5′-GGTAC/C-3′ to generate DNA fragments with 3′-cohesive termini.