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. 2018 Jul 30;8:11392. doi: 10.1038/s41598-018-29621-1

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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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Assay sensitivity and anomalous DL copy number. (A) DL2:BA ratio from multiplex assay of gDNA from A7r5 cells grown in normal culture media without (CTRL) or with 2,3-dideoxycitidine (ddC, 30 µM) for 2–14 days. Error bars are standard error. (B) Assay sensitivity was assessed by titrating gDNA from control and ddC-treated cells to create known levels of mtDNA depletion. *All pair-wise comparisons are significantly different other than for the 5% and 10% groups. C. Singleplex droplet distributions for ND and DL2 primers amplifying A7r5 gDNA. (D) Copy numbers from C normalized to ND5 for each experiment. (E) Representative singleplex droplet distributions for DL2 and flanking primers (see Fig. 1) using A7r5 gDNA. (F) Copy numbers from E normalized to NCR1. Error bars are 95% confidence interval of Poisson-corrected concentrations (G) Typical droplet intensity distribution for the ND1-DL2 assay. (H) Sister cultures of A7r5 cells treated with ddC (as in A) or vehicle (control), and analyzed for DL2:ND1 and ND3:ND1 ratios. Dotted line indicates no difference. Data points below line show depletion by ddC.