Measuring Cation Dependent DNA Polymerase Fidelity Landscapes by Deep Sequencing
(A) Overview of a strategy for using DNA polymerases as signal recording devices. Signals (top) are coupled to intracellular or extracellular cation concentration through direct or indirect modulation of an ion channel activity. Cation concentration is in turn coupled to DNA polymerase fidelity on a known template according to a known transfer function (orange curve), generating a DNA recording, in which data is represented by the density of misincorporated bases, and which can be read by DNA sequencing (bottom). (B) Modulation of Taq polymerase by Ca2+ concentration, measured by a traditional blue-white colony counting assay . (C) Biochemical steps of the multiplex deep sequencing assay for measuring the transfer functions of error-prone DNAPs.