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Nco I
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Nco I Technical BrochureTechnical Brochure
NcoI is a restriction enzyme purified from Nocardia corallina.

 

Unit substrate: Lambda DNA.

 

Unit calculation assay conditions: 100 mM NaCl, 50 mM Tris-HCl (pH 7.9 @ 25oC), 10 mM MgCl2, 1 mM DTT, 0.02% Triton X-100, 100 μg/ml bovine serum albumin and DNA. Incubate at 37oC.

 

Absence of contaminants: 100 units of Nco I do not produce any unspecific cleavage products after 16 hrs incubation with 1 μg of Lambda DNA at 37oC. After 50-fold overdigestion with Nco I, 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: 65oC for 20 minutes.

 

Methylation Sensitivity:

dam methylation: Not sensitive

dcm methylation: Not sensitive

CpG methylation: Not sensitive

 

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

 

Reagents supplied: 10x UNcoI and 10x K buffer