TA Cloning - old product

hodges bradley l segdoh at uxa.cso.uiuc.edu
Mon Feb 6 15:47:29 EST 1995


Did you remove the terminal phosphates from your TA vector prior to 
ligation?  This dramatically improves efficiency.X

On Wed, 1 Feb 1995, Darren M. Stauth wrote:

> In article <D3B0Kr.JwG at zoo.toronto.edu>, mes at zoo.toronto.edu (Mark Siddall)
> wrote:
> 
> > 
> > 
> > I am attempting to clone my pcr product with the TA system.  First
> > attempt failed.
> > Product was about a week old but had been at -20C the whole time.
> > Now it's about 2 weeks old though still at -20C.
> > Any body know what the likelihood of having lost the adenosine
> > residues might be?
> > 
> > Mark
> > 
> > -- 
> > Mark E. Siddall                "I don't mind a parasite...
> > mes at vims.edu                    I object to a cut-rate one" 
> > Virginia Inst. Marine Sci.                     - Rick
> > Gloucester Point, VA, 23062
> 
> Protocols suggest you use fresh product.  However my last TA cloning of
> about 22 different samples was done using PCR products at least 3 weeks old
> and it still worked.  I do think the results would have been better if I
> had used fresh template.
> 
> 



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