Artifactual mutations resulting from DNA lesions limit detection levels in ultrasensitive sequencing applications.

ABSTRACT: The need in cancer research or evolutionary biology to detect rare mutations or variants present at very low frequencies (<10^-5) poses an increasing demand on lowering the detection limits of available methods. Here we demonstrated that amplifiable DNA lesions introduce important error sources in ultrasensitive technologies such as single molecule PCR (smPCR) applications (e.g. droplet-digital PCR), or next-generation sequencing (NGS) based methods. Using templates with known amplifiable lesions (8-oxoguanine, deaminated 5-methylcytosine, uracil, and DNA heteroduplexes), we assessed with smPCR and duplex sequencing that templates with these lesions were amplified very efficiently by proofreading polymerases (except uracil), leading to G->T, and to a lesser extent, to unreported

SUBMITTER: Arbeithuber B 

PROVIDER: S-EPMC5144678 | biostudies-literature | 2016 Dec

REPOSITORIES: biostudies-literature