>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
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.