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. 2017 Aug 9;7:7683. doi: 10.1038/s41598-017-08067-x

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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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Analysis of the reaction kinetics with respect to different hinge-primer combinations using dSpacer as blocking-site. (a) Overview of primer combinations for symmetric reaction (1–2) and asymmetric reaction (3–7) (b) Normalised real-time amplification curves for each combination. (c) Melting-curve analysis. Red: combination 1, green: combination 2, blue: combination 4, grey: combination 3, brown: combination 6, violet: combination 5 and black: combination 7. Solid lines represent reactions with 5 ng genomic DNA (E.coli) and dashed lines are non-template controls.