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TA cloning vectors

berschic at convex.rz.uni-duesseldorf.de berschic at convex.rz.uni-duesseldorf.de
Tue Jul 5 08:32:16 EST 1994

>Dear Herb and Peter and others who are interested,
>I have a ref for constructing T-vectors (Marchuk et al, NAR, 19 no 5,
>p1154). It uses dTTP alone with taq polymerase, which results in a single
>thymidine at the 3' end of the vector. (If you use ddTTP you won't get
>ligation to your PCR product unless you add phosphates to the end of the
>You can use this principle to make blunt end ligations more efficient -
>incubate insert with taq and dATP, vector with taq and dTTP.
>I'm trying it right now, so i hope it works
>Mill Hill, London, UK
Hi Melanie,
I found the citation for the ddTTP vectors for cloning. It is :
Holton et al., 1991: A simple method for direct cloning of PCR-products 
using ddT-tailed vectors. NAR 19:1156.

I didn´t use it but why shouldn´t it work? The ligation of 
dephosphorylated vector (to inhibit self ligation) with PCR product does
also work. The 5´end of vector could ligate with 3´ end of PCR-product. 
Perhaps the not ligated 3' end of vector (ddT) could be repaired by 
excision and replacing of ddT during replication in the host. 

Uni Duesseldorf
Duesseldorf, Germany

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