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)
> > 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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