Abstract

We tested the ability of Taq DNA polymerase (Taq) to amplify long DNA fragments and showed that, if the conditions were set properly, Taq could successfully perform the “long PCR” up to 24 kb. The conditions include: (1) longer primers, (2) a 2-step cycling, and (3) a “long buffer.” We propose that the most important requirements are the survival rate of Taq at high temperatures and that of the primers against the 5 to 3 exonuclease activity of Taq.

Keywords: Taq DNA polymerase, long amplicons, primers, reaction buffer