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. 2018 Feb 1;8:2028. doi: 10.1038/s41598-018-19501-z

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© The Author(s) 2018

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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MIP protocol optimization for the 16S probeset. Pooled 16S MIPs were tested against DNA extracts of representative biothreats and ESKAPE pathogens to optimize target capture conditions including (a) probe concentration, (b) reaction temperature and (c) reaction duration. Amplicon formation was quantitatively measured after probe circularization and amplification with the universal primer set across three replicates for each organism at each variable. Concentrations were normalized to 0% and 100% representing the lowest and highest concentrations across the experimental range. Data points for each organism were combined and a corresponding outlier box plot was generated for each variable.