Nru I digestion problems

Karl Fischer tyr-2 at bones.biochem.ualberta.ca
Wed Feb 21 17:52:58 EST 1996


Hello,

We are having problems digesting one of our plasmids with the restriction
enzyme Nru I. The plasmid was propogated in E. coli DH5alpha, purified on
a Quiagen column, and evaluated by UV spec (260/280 ratio 1.9). Five
micrograms was digested O/N at 37 degrees with 20 U in a final volume of
20 ul then qs'd to 40 ul and further digested with an additional 20U of
enzyme. Gel analysis shows no digestion.

Has anyone experienced wonky cutting with this enzyme? We are currently
sequencing the region where the Nru site lies, though the DNA containing
the site is from a prototype clone (ie. fully sequenced by another
laboratory).

Any information is greatly appreciated.

TIA

Karl the hepB guy

-- 
Karl Fischer
tyr-2 at bones.biochem.ualberta.ca





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