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. 2017 Jul 5;7:4688. doi: 10.1038/s41598-017-04931-y

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Validation of the next-generation sequencing assay. (A) A position-specific error rate per nucleotide was determined before or after read-cleaning algorithms were applied for the control experiments with plasmid (P) or RNA transcript (T) in 3 independent next-generation sequencing experiments (1–3) each. Mean ± sem are shown. (B) For confirmation, a position-specific error rate per amino acid position was determined after read-cleaning algorithms were applied for the control experiments with plasmid (P) or RNA transcript (T) in 3 independent next-generation sequencing experiments (1–3) each. Mean ± sem are shown. (C) Haplotypes and their frequencies in the dataset after read-cleaning were determined for the control experiments with plasmid (P) or RNA transcript (T) in 3 independent next-generation sequencing experiments (1–3) and plotted against each other with a cut-off value of 0.1%.