Home-Made TA cloning

Shane Albright insomnia at mendel.berkeley.edu
Tue Jul 18 20:33:28 EST 1995


In article <m-kamei-1807950946480001 at mc-ii2.waite.adelaide.edu.au>,
m-kamei at schooner.waite.adelaide.edu.au (Makoto Kamei) wrote:

> >Surely that should be ddTTP. If you use just dTTP then terminal transferase
> >will add umpteen 'T's.
> 
> It is dTTP. I've done this and it worked quite well. My superviser was
> boasting that this works better than pGEM-T cloning system from Promega.
> She was saying that bluescript works better than pGEM for colour
> selection.

Yep, it's dTTP.  Taq's "terminal transferase activity" only adds a single
3' base.  And this homemade version definitely works better than Promega
or Novagen vectors.

-- 
Shane Albright
MCB Dept, UC Berkeley
insomnia at mendel.berkeley.edu



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