TA Cloning Vectors

[pzkok at FLINDERS.EDU.AU]

>Hi, an old question to this group.  What TA cloning vector have
>individuals used sucessfully for cloning PCR amplified cDNA fragments of
>approximately 600-800 bp.  I have heard that Invitrogen vector is good,
>but unstable.  Has anyone used the new Genehunter TA vector?
>
>Any suggestions will be appreciated.
>
>
We make our own TA cloning vector by using any cloning vector (preferably with 
blue-white selection) with an EcoRV-site. Cleave it with EcoRV in Taq-buffer, 
inactivate and incubate with only dTTP and Taq-polymerase 2hrs 72 degrees. 
Purify with your favourite method and use for ligation. This works for us, we 
get about 50% positives of the white colonies fron a blue-white plating.
This method was published in NAR 1992 or 1993 but I don't remember the exact 
reference, sorry.

Good Luck
Klas