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Kpn I
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Kpn I Technical BrochureTechnical Brochure
KpnI is a restriction enzyme purified from Klebsiella pneumonia OK8.

 

Unit substrate: Lambda DNA (EcoRI digest).

 

Unit calculation assay conditions: 10 mM Tris-HCl (pH 7.0 @ 25oC), 10 mM MgCl2, 1 mM dithiothreitol, 0.01% Triton X-100, 100 μg/ml bovine serum albumin and DNA. Incubate at 37oC.

 

Absence of contaminants: 30 units of Kpn I do not produce any unspecific cleavage products after 16 hrs incubation with 1 μg of lambda DNA/EcoR I digest at 37oC. After 10-fold overdigestion with KpnI, greater than 95% of the DNA fragments can be ligated and recut with this enzyme.

 

Storage buffer: 50 mM KCl, 10 mM Tris-HCl (pH 7.4), 0.1 mM EDTA, 1 mM dithiothreitol, 200 μg/ml bovine serum albumin and 50% glycerol. Store at -20oC.

 

Heat inactivation: No.

 

Methylation Sensitivity:

dam methylation: Not sensitive

dcm methylation: Not sensitive

CpG methylation: Not sensitive



Star activity: Conditions of low ionic strength, high enzyme concentration, glycerol concentration >5% or pH >8.0 may result in star activity.

 

Percent Activity in MINOTECH Buffers
 L M  H  SH  A  K 
75-100  25-50  <10  <10  50  100 
 
 
General reaction mixture:
10U KpnI 1μl
10x UKpnI or K buffer*  2μl 
DNA substrate  <1μg
Sterile ultrapure water  Up to 20 μl
Incubate for 15 min at 37oC  
*In the case of UKpnI buffer we recommend the addition of BSA to a final concentration of 100 μg/ml. 

 

Reagents supplied: 10x UKpnI and 10x K buffer