Engineered DNA Polymerases Improve NGS and Enable Novel Applications

The vast majority of PCR applications utilize a small number of wild-type DNA polymerases; improvements to existing applications and the development of novel PCR applications have primarily been realized via innovative changes to all of the reaction components except the polymerases themselves. More recently, KAPA have engineered a new generation of improved DNA polymerases for PCR applications, including KAPA HiFi, a high-fidelity, proof-reading DNA polymerase. KAPA HiFi is capable of robust amplification of diverse targets spanning a wide range of sizes and GC-content, and KAPA have demonstrated that KAPA HiFi produces high yields with remarkably little amplification bias when used for NGS library amplification. With improved Methyl-seq performance in mind, KAPA further engineered KAPA HiFi to allow the efficient amplification of bisulfite-converted DNA libraries containing uracil. Using a number of case studies, it is illustrated that these engineered DNA polymerases are providing better sequence coverage and are enabling novel and challenging NGS applications.