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. 2021 Jan 21;22:69. doi: 10.1186/s12864-021-07381-z

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© The Author(s) 2021

Open AccessThis 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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 6 — Freeze-Thaw Cycles Exacerbate 3′ Bias in poly(A)-enriched samples. a Gene coverage (y-axis) at the i^th nucleotide percentile (x-axis) for samples that underwent 1–5 freeze-thaws and were extracted using either poly(A)-enrichment or ribosomal depletion. Coverage is normalized to samples that underwent one freeze-thaw. For each sample, coverage is averaged across all genes; samples are aggregated using generalized additive model smoothing, with shaded regions representing 95% confidence intervals. b Linear model fits comparing the change in normalized median coverage percentile to the number of freeze-thaw cycles (log-transformed) for ribosomal depletion (orange) or poly(A)-enrichment (green)