TA Cloning: a skeptic

[Michael Cooley]

Tanya Reichert (reichert at bio.bu.edu) wrote:
: This may be a really stupid question but...Why is the TA vector (in whatever
: incarnation - Invitrogen, own making) unstable?  If you resuspend it in 
: clean, nuclease-free water or buffer, how does it self-destruct?  One view
: I read was that the T's come off.  How is that possible?  Does the phospho-
: diester bond hydrolyze spontaneously?  Am I totally off my rocker in being 
: a skeptic of this degradation / instability?  Or is this some marketing 
: gimmick of the manufacturer?

: I guess the bottom line is that I'm having a bloody awful time trying to 
: subclone my PCR products and all this frustration is making me grouchy.


: May the PCR and cloning gods smile upon all of us.


: Tanya


An Invitrogen rep told me the company was experimenting with water 
sources to improve the stability of the vector. My experience indicates 
that really old vector worked badly. 


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