Polymerase Specific Error Rates and Profiles Identified by PacBio Single Molecule Sequencing. Polymerase Error Rates by PacBio Sequencing

DNA polymerases have an innate error rate which is polymerase and DNA context specific. We used the unique properties of PacBio single molecule sequencing to directly evaluate the mutational rate and profiles of amplicons generated by six commonly used DNA polymerases. Primers targeted the CASK and FAM120C genes and included barcodes and padding sequence from PacBio's initial 48 barcode design. The following barcodes and polymerases were used: 1-Phusion High-Fidelity DNA Polymerase, 2-KAPA HiFi DNA Polymerase, 3-HotStar HiFidelity Polymerase Kit (Qiagen), 4-TaKaRa LA Taq DNA Polymerase, 5-Platinum Taq DNA Polymerase (Invitrogen), 6- Illumina’s Nano kit polymerase. This enabled error analysis by nucleotide error counts and direction specific mutations through the analysis of heteroduplexes. Error rates, as well as the most common transitions, differed by polymerase.