Separating cut from uncut vector

John K. Troyer jtroyer at
Wed Aug 23 08:12:56 EST 1995

On 23 Aug 1995, Ira M. Lubin wrote:

> I'm looking for suggestions on how to effectively separate a 12kb  vector 
> fragment from uncut vector.  0.7%-1.0% agarose gel
> electrophoresis is not sufficient. Enough uncut vector remains to 
> adversely effect subsequent ligation/transformation.
> Thanks,
> Ira

Non-denaturining acrylamide gels (5-10%) give MUCH better separation than 
agarose gels.  The vector can be separated and eluted from the gel in much the 
same way as you do with agarose. 


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