TA cloning

rdudley at my-dejanews.com rdudley at my-dejanews.com
Thu Jun 18 10:26:07 EST 1998


In article <6m8ss3$cog at mserv1.dl.ac.uk>,
  Denni Schnapp <dschnapp at molbiol.ox.ac.uk> wrote:
>
>
> Hi,
>
> I want to TA clone some PCR products, but these were kept at -20 C for a
> few weeks. Would this cause problems? Are there better ways (if any) of
> storing the PCR reactions? Also, what is people's experience with kits
> (TA, not blunt-end, and preferably cheap)? I am thinking about getting
> PGEM T vector from Promega, seem to be the cheapest in Europe.
>
> Regards,
>
> Denni
>
> Dept. Paediatric Gastroenterology
> Bart's Hospital
> Dominion House
> 59 St. Bartholomews Close
> London EC1A 7BE

I have had mediocre success with PCR products frozen at -20C for even a few
days.  I recommend using fresh products, if that's not too much of a bother.
You could try and +re-tail+ heproducts by incubating at 72C (if Taq) with dATP
for about 10 min.

As for the kit, I swear by Invitrogen's Original TA cloning kit.However, it is
the onlyone I have ever tried.

rich

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