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Nru I
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Nru I Technical BrochureTechnical Brochure
NruI is a restriction enzyme purified from Nocardia rubra.

 

Unit substrate: Lambda DNA.

 

Unit calculation assay conditions: 100 mM KCl, 50 mM Tris-HCl (pH 8.0 @ 25oC), 10 mM MgCl2, 100 μg/ml bovine serum albumin and DNA. Incubate at 37oC.

 

Absence of contaminants: 80 units of Nru I do not produce any unspecific cleavage products after 16 hrs incubation with 1 μg of Lambda DNA at 37oC. After 10-fold overdigestion with Nru I, less than 20% of the DNA fragments can be ligated.

 

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: Blocked by overlapping

dcm methylation: Not sensitive

CpG methylation: Blocked

 

Star activity: Large excess of the enzyme results in the appearance of star activity.

 

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

 

Reagents supplied: 10x UNruI and 10x K buffer