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. 2020 Jul 7;10:11147. doi: 10.1038/s41598-020-67825-6

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© The Author(s) 2020

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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The figure depicts three exemplary amplification plots (unamplified, 10 × and 14 × pre-amplified) from one saliva sample (sample ID 7) and one gene (CDKN1A) created with the ABI PRISM 7900HT sequence detection system. The amplification plot (A) represents the unamplified sample, the one marked with (B) the 10 × pre-amplified sample and the one with (C) the 14 × pre-amplified sample. Droplines (vertical dashed lines) provide the corresponding cycles required for the fluorescent signal to cross the threshold (Ct values). Linearity of 10 × pre-amplification is shown as a ΔCt of 10 in horizontal solid line 1 (Ct _(no-Amp)—Ct _{(14X pre-Amp)}), and corresponding ΔCt of 4 in horizontal dotted line 2 (Ct _{(10X pre-Amp)}—Ct _{(14X pre-Amp)}). ΔRn (normalized reporter value): the Rn value of an experimental reaction minus the Rn value of the baseline signal.