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. 2022 Jul 13;21:139. doi: 10.1186/s12934-022-01844-y

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© The Author(s) 2022

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. 5 — Effect of the terminator sequences on mutagenesis efficiency. A Average mutagenesis efficiency expressed as the percentage of colonies identified as mutants by colony PCR genotyping for every terminator sequence tested in three independent biological replicates. The colour code (scale bar shown on the right side of panel B represents the termination strength as determined by Chen et al. [11]. The statistical analysis shows the results of a one-way ANOVA followed by a Holm-Sidak’s multiple comparisons test comparing all values to the control sample (deletion oligonucleotide). P values: P ≤ 0.001 (***), P ≤ 0.01 (**), P ≤ 0.05 (*), P ≤ 0.1 (0.1), N.S. (not significant). B Average mutagenesis efficiency represented as colour-coded percentage of colonies identified as mutants by colony PCR genotyping in correlation to the ΔGfree of the secondary structure formation of the terminator sequences as calculated by the Vienna RNA suite [27]. The curved dotted solid lines represent the 95% confidence interval of the linear regression (solid line). P value of R²: P ≤ 0.01 (**)