|
Download
article
229kb
pdf? |
|
Cloning Blunt-End DNA Fragments Into the pGEM®-T Vector Systems
By Gary Kobs
Promega Corporation
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.
|
