High-resolution mapping of DNA polymerase fidelity using nucleotide imbalances and next-generation sequencing.

ABSTRACT: DNA polymerase fidelity is affected by both intrinsic properties and environmental conditions. Current strategies for measuring DNA polymerase error rate in vitro are constrained by low error subtype sensitivity, poor scalability, and lack of flexibility in types of sequence contexts that can be tested. We have developed the Magnification via Nucleotide Imbalance Fidelity (MagNIFi) assay, a scalable next-generation sequencing assay that uses a biased deoxynucleotide pool to quantitatively shift error rates into a range where errors are frequent and hence measurement is robust, while still allowing for accurate mapping to error rates under typical conditions. This assay is compatible with a wide range of fidelity-modulating conditions, and enables high-throughput analysis of sequence contex

SUBMITTER: de Paz AM 

PROVIDER: S-EPMC6061839 | biostudies-literature | 2018 Jul

REPOSITORIES: biostudies-literature