Home-Made TA cloning

[Daniel Kim]

In article <Stephen_Lasky-1007951408300001 at tonto-slip2.cis.brown.edu> Stephen_Lasky at brown.edu (Stephen R. Lasky, Ph.D.) writes:
> > 
[omitted]
>> 
>> 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.
>

You are correct.  Taq will add one dNTP onto the end of a double stranded 
DNA, but will not add onto the end of a ssDNA, so the overhang is not a 
substrate for further extension.

Daniel Kim