ArticleScientific reports2017
A high-throughput assay for quantitative measurement of PCR errors.
Article in Scientific reports, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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Who cites it
15 citing papers in PubMed.
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- Next-Generation Sequencing Methods to Determine the Accuracy of Retroviral Reverse Transcriptases: Advantages and Limitations.Viruses · 2025Review
- Correcting PCR amplification errors in unique molecular identifiers to generate accurate numbers of sequencing molecules.Nature methods · 2024Article
- Long amplicon nanopore sequencing ofBiology methods & protocols · 2024Article
- A digital twin for DNA data storage based on comprehensive quantification of errors and biases.Nature communications · 2023Article
- Liquid Biopsies in Lung Cancer.Cancers · 2023Review
- Phasing of de novo mutations using a scaled-up multiple amplicon long-read sequencing approach.Human mutation · 2022Article
- Prognostic Value of Circulating Tumor DNA (ctDNA) in Oncogene-Driven NSCLC: Current Knowledge and Future Perspectives.Cancers · 2022Review
- Genotypic Characterization of ClinicalFrontiers in microbiology · 2021Article
- A novel virtual barcode strategy for accurate panel-wide variant calling in circulating tumor DNA.BMC bioinformatics · 2020Article
- TILLING by Sequencing: A Successful Approach to Identify Rare Alleles in Soybean Populations.Genes · 2019Article
- Deep sequencing of B cell receptor repertoire.BMB reports · 2019Article
- smCounter2: an accurate low-frequency variant caller for targeted sequencing data with unique molecular identifiers.Bioinformatics (Oxford, England) · 2019Article
- A droplet microfluidic platform for efficient enzymatic chromatin digestion enables robust determination of nucleosome positioning.Lab on a chip · 2018Article
- MERIT reveals the impact of genomic context on sequencing error rate in ultra-deep applications.BMC bioinformatics · 2018Article
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Authors and funding
8 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The accuracy with which DNA polymerase can replicate a template DNA sequence is an extremely important property that can vary by an order of magnitude from one enzyme to another. The rate of nucleotide misincorporation is shaped by multiple factors, including PCR conditions and proofreading capabilities, and proper assessment of polymerase error rate is essential for a wide range of sensitive PCR-based assays. In this paper, we describe a method for studying polymerase errors with exceptional resolution, which combines unique molecular identifier tagging and high-throughput sequencing. Our protocol is less laborious than commonly-used methods, and is also scalable, robust and accurate. In a series of nine PCR assays, we have measured a range of polymerase accuracies that is in line with previous observations. However, we were also able to comprehensively describe individual errors introduced by each polymerase after either 20 PCR cycles or a linear amplification, revealing specific substitution preferences and the diversity of PCR error frequency profiles. We also demonstrate that the detected high-frequency PCR errors are highly recurrent and that the position in the template sequence and polymerase-specific substitution preferences are among the major factors influencing the observed PCR error rate.
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