Home-Made TA cloning

Duncan Clark Duncan at genesys.demon.co.uk
Mon Jul 10 07:10:12 EST 1995


In article: <3tps79$chq at agate.berkeley.edu>  Tjian lab <insomnia at mendel.berkeley.edu> writes:
> 
> s060965 at AIX1.UOTTAWA.CA wrote:
> >I've been trying to think of a way to make my own TA cloning kit.  
> >
> {Stuff deleted}
> 
> >Any ideas out there.
> >
> >Cris Martin
> 
> 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
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