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. 2017 Oct 17;7:13394. doi: 10.1038/s41598-017-13881-4

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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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Reaction results based on different combinations of reaction components according to an orthogonal test. (a) Reactions of 16 combinations monitored by the amplification curves. (b) Corresponding melt peaks of the above reactions. 1–16 represent each combination performed in the orthogonal test shown in Table 1. NC indicates reactions that share the same conditions with each combination except for the substitution of a negative control template (genomic DNA of Sporisorium reiliana).