Directed evolution of DNA polymerases for next-generation sequencing

Abstract

We present the application of an activity-based phage display method to identify DNA polymerases tailored for next generation sequencing applications. Using this approach, we identify a mutant of Taq DNA polymerase that incorporates the fluorophore-labeled dA, dT, dC, and dG substrates ~50 to 400-fold more efficiently into scarred primers in solution and that also demonstrates significantly improved performance under actual sequencing conditions.