Cloning Blunt-End DNA Fragments Into the pGEM®-T Vector Systems

Blunt-end DNA fragments can be ligated into Promega's T-Vectors if they are first "tailed" using dATP and Taq DNA Polymerase. In this article, the results of two different A-tailing protocols were evaluated using a pGEM®-T Easy Vector System. A standard A-tailing protocol was compared to an abbreviated alternative protocol using both PCR products generated by thermostable DNA polymerases that have proofreading activity, and modified blunt-end DNA fragments generated by restriction digestion. The efficiency of cloning long PCR fragments into the pGEM®-T Vector Systems was also evaluated.

Promega Notes 62, 15.