Construction of TA cloning vector

Helge Weissig helgew at
Mon Jan 31 20:17:06 EST 1994

cdolphin at (Dr. C. Dolphin) wrote:

>you could also incubate blunt-cut vector with dideoxyTTP (ddTTP) and terminal
>transferase which will add single ddTTP to each 3'end. Addition of extra dATP
>in the final elongation stage of PCR may also increase proportion of products
>with A overhangs (question - does Taq add just a single dATP or does it also
>have terminal transferase-like activity?). We have used this for succssefully
>for sub-cloning of many PCR products. Also include a touch of the enzyme used
>to blunt cut vector in the ligation reaction to reduce background.


how come that you get ligation to a ddTTP tailed DNA strand?



Helge Weissig      

La Jolla Cancer Research Foundation             (619) 455 6480 x253
10901 N. Torrey Pines Rd.                  FAX: (619) 455 0181
La Jolla, CA 92034                      e-mail: helgew at


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