Polymerase specific error rates and profiles identified by single molecule sequencing

•PacBio single molecule circular sequencing enables the determination of polymerase error rates and profiles.•Low-fidelity and high fidelity polymerases demonstrate approximately ten fold difference in error rates and can be discriminated by error profiles.•PacBio sequencing of heteroduplexes enables the determination of direction specific transversions and transitions.•Watson–Crick base pairing errors are not equally distributed, with pyrimidine transitions occurring more frequently than purine transitions.

DNA polymerases have an innate error rate which is polymerase and DNA context specific. Historically the mutational rate and profiles have been measured using a variety of methods, each with their own technical limitations. Here we used the unique properties of single molecule sequencing to evaluate the mutational rate and profiles of six DNA polymerases at the sequence level. In addition to accurately determining mutations in double strands, single molecule sequencing also captures direction specific transversions and transitions through the analysis of heteroduplexes. Not only did the error rates vary, but also the direction specific transitions differed among polymerases.