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