TA cloning kit

[David Micklem]

In article <38284ced.0 at news.uni-ulm.de>, Dr. med. Franz F. Wagner
<fwagner at medizin.uni-ulm.de> wrote:

>For several years, we used the Amersham pMosBlue T-vector kit for TA
>cloning. It was very satisfying with normal Taq and with Boehringer
>high-fidelity PCR system. Now I had to learn that Amersham do no more
>produce this kit and instead sell a blunt-end cloning kit that they claim to
>be even superior.
>
>My questions:
>Is this blunt-end pMosBlue PCR cloning kit really comparable or even better
>to the pMosBlue T-vector kit (my insert size is about 500 to 5,000 bp)?
>Are there other companies selling _good_ T-vector kits?
>
>I would be happy for any recommendations based on personal experience.
>
>I thank you very much in advance

Notwithstanding recent negative experiences reported on this newsgroup
(with rearranged clones/loss of polylinker sequences) I have found the
Invitrogen TOPO-TA vectors to work fantastically well.

They are kind of expensive as they insist on supplying competent cells
with the vector, but if you have  a source of other cells you can make
the vector last at least 2-3 times as long by scaling down their
recommended reaction.

David

-- 
D.R. Micklem,          Time flies like an arrow... Fruit flies like a banana. 
Beckman Center,        Email:dmicklem at cmgm.stanford.edu
Stanford University    Phone: +1 (650) 723-6650
Stanford, Ca 94305       Fax: +1 (650) 725-6044
USA                    Unsolicited email will incur a US$100 processing charge.