TA vs topoTA cloning

rdudley at my-dejanews.com rdudley at my-dejanews.com
Fri Jun 19 09:20:58 EST 1998


In article <1998061901051400.VAA08651 at ladder01.news.aol.com>,
  greggsilk at aol.com (Gregg Silk) wrote:

<snip>
>
> I have used the original InVitrogen TA and the new topoTA kit. Both of them
> worked well, and the 5 min "ligation" of the topo kit is slick. The blue/white
> screen for the topo kit is strong, although there are plenty of blues. And
> although the topo kit uses a special reagent, they provide enough for 20 or so
> reactions.
> Gregg
>

One consideration for TOPO-TA cloning (and this really should hold true for
*everything* that is done in the lab)--make sure you read the background
literature as to how topoisomerases work!  I've mentioned this before in this
newsgroup--our cDNA contains a recognition sequence for topoisomerases.  This
ended up supercoiling our starting material, which given the design of the
project, was a horrendous pain to deal with.

rich

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