Home-Made TA cloning

Stephen R. Lasky, Ph.D. Stephen_Lasky at brown.edu
Mon Jul 10 14:08:30 EST 1995


 > 
> > Actually, it's a lot easier than that.  Just linearize bluescript with 
> > EcoRV and incubate with Taq and dTTP at 72 deg for 2 hrs.  The terminal 
> > transferase activity is non-specific and will use dTTP if nothing else 
> > is available.  See Marchuk et al, NAR 19(5):1154 for details
> 
> Surely that should be ddTTP. If you use just dTTP then terminal transferase
> will add umpteen 'T's.
> 
Duncan:  I think that it IS dTTP.  The terminal transferase that the
previous writer is speaking of is the activity associated with Taq Poly,
and, from my understanding, that ususally just adds a single non-template
driven base.  If I'm wrong, I'd appreciate knowing.

SRLasky

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Stephen R. Lasky Ph.D.  Brown University/Roger Williams Medical Center
Landline: 401-456-6572   Fax: 401-456-6569  E-Mail: Stephen_Lasky at brown.edu
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