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. 2024 Jan 8;14:828. doi: 10.1038/s41598-023-50667-3

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© The Author(s) 2024

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Pacific Biosciences (PacBio) single-molecule sequencing traces of the C9orf72 repeat region for patient and control cell lines. (A–H) Sequencing traces showing the number of circular consensus sequencing (CCS) reads per repeat count. In the sequencing traces, each horizontal line represents one sequenced molecule of DNA. Blue color depicts on-target sequencing with GGGGCC repeat, grey color depicts sequencing error. Each molecule is anchored to an adjacent, non-repeat region (CGCCC) which is not included In the total repeat count. Y-axis = CCS count. In the WT line, repeat length has a bimodal distribution with roughly equal numbers of reads containing 2 or 10 repeats, indicating that one allele has 2 repeats and the other 10. In the patient lines, a bimodal distribution is present but not always as apparent: all lines show a peak with a low repeat number (2–10), corresponding to the unexpanded allele, and the expanded allele size can vary across cell lines from different donors.