Help: How do I get rid of EDTA and T4kinase from my oligos

Bill Alexander alexanderw at cber.cber.fda.gov
Tue Oct 15 09:37:39 EST 1996


In Article <randyg-0910961048440001 at crl228-200-218.crl.umn.edu>,
randyg at puccini.crl.umn.edu (Dr. Randal W. Giroux) wrote:
>We are adding some linkers to our vector to change the restriction site
>and we have to phosphorylate one of the two oligos (16mer) before we
>ligate them to the vector.
>
> BUT once we finish the reaction with EDTA we need to use these oligos in
>the next ligation reaction. The t4 kinase enzyme and EDTA are still around
>and the EDTA will inhibit ligation and the T4 kinase will still be active.
>How can I purify the oligos away from the phosphorylation reaction
>components. Are there kits or quick protocols to do this. I'm stumped
>
Is this a troll for a response Dr?  Stumped?  Really?

Try Maniatis et al., (Molecular Cloning: A Laboratory Manual, any edition),
or any other Molecular Biology Lab Manual.  Many company catalogs will even
tell you how to do this.   

Bill Alexander  



More information about the Methods mailing list