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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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Single factor experiment of RealAmp. The influence of each variable on the LAMP reaction was analyzed by the amplification curve (a) and the melt peak (b). For the Bst DNA polymerase optimization, Bst polymerase quantities were adjusted to 2 U, 4 U, 6 U and 8 U, respectively. Ratios of inner to outer primers were set at 2:1, 4:1, 6:1 and 8:1 with the outer primer concentration fixed to 0.2 μM to optimize the primer ratios. Mg²⁺ concentrations in the LAMP reactions were varied from 5 mM, to 8 mM for the optimization of Mg²⁺.