Figure 1. Scheme of a phosphorothioated-terminal hairpin formation and self-priming extension (PS-THSP) for selection ... Figure 1. Scheme of a phosphorothioated-terminal.

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Figure 1. Scheme of a phosphorothioated-terminal hairpin formation and self-priming extension (PS-THSP) for selection ... Figure 1. Scheme of a phosphorothioated-terminal hairpin formation and self-priming extension (PS-THSP) for selection of self-replicators. Unless provided in the caption above, the following copyright applies to the content of this slide: © The Author(s) 2019. Published by Oxford University Press on behalf of Nucleic Acids Research.This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com Nucleic Acids Res, Volume 47, Issue 5, 30 January 2019, Pages 2169–2176, https://doi.org/10.1093/nar/gkz044 The content of this slide may be subject to copyright: please see the slide notes for details.

Figure 5. Real-time data of PS-THSP for high and low ranked R region sequences. (A) Two HH and two LL sequences whose ... Figure 5. Real-time data of PS-THSP for high and low ranked R region sequences. (A) Two HH and two LL sequences whose ranks remained high and low over four rounds were selected, respectively. (B) PS-THSP reactions were monitored in a real-time using a SYTO 82 dye with different concentrations of Bst DNA polymerase. Unless provided in the caption above, the following copyright applies to the content of this slide: © The Author(s) 2019. Published by Oxford University Press on behalf of Nucleic Acids Research.This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com Nucleic Acids Res, Volume 47, Issue 5, 30 January 2019, Pages 2169–2176, https://doi.org/10.1093/nar/gkz044 The content of this slide may be subject to copyright: please see the slide notes for details.

Figure 4. Analysis of different probe 1 replicators at individual positions in the R region during the first round of ... Figure 4. Analysis of different probe 1 replicators at individual positions in the R region during the first round of selection. (A) A heat map was prepared to show the relative frequency of each base (A, G, T, C) at each position within top ranking sequences (top 100, 1K, 10K, 50K, 100K, 200K, and 500K). Frequency trends for dinucleotide (B) and trinucleotide (C) sequences within top ranking 100 and 500K sequences were again represented using heat maps. (D) The top 10 ranked sequences from the first round for probe 1. Unless provided in the caption above, the following copyright applies to the content of this slide: © The Author(s) 2019. Published by Oxford University Press on behalf of Nucleic Acids Research.This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com Nucleic Acids Res, Volume 47, Issue 5, 30 January 2019, Pages 2169–2176, https://doi.org/10.1093/nar/gkz044 The content of this slide may be subject to copyright: please see the slide notes for details.

Figure 3. Evolution of selected R region sequences Figure 3. Evolution of selected R region sequences. (A) Rankings of the top 500 sequences from the first through fourth ... Figure 3. Evolution of selected R region sequences. (A) Rankings of the top 500 sequences from the first through fourth rounds of selection for PS-THSP probe 1. (B) Sequence logos for the top 100 PS-THSP probe 1 and probe 2 sequences over four rounds. (C) The yellow bars in the graph represent the identity of the initial nucleotide for the top 500 ranked sequences in each of the 4 rounds of selection, with the y axis showing the rank of the sequences (higher numbers of sequences towards the top). (D) Scheme for extension without concatamerization of the PS-THSP template, and corresponding analysis of the resultant R region sequences at different positions for PS-THSP probe 1 and probe 2. The x and y axes represent base position and count of sequences, respectively. Unless provided in the caption above, the following copyright applies to the content of this slide: © The Author(s) 2019. Published by Oxford University Press on behalf of Nucleic Acids Research.This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com Nucleic Acids Res, Volume 47, Issue 5, 30 January 2019, Pages 2169–2176, https://doi.org/10.1093/nar/gkz044 The content of this slide may be subject to copyright: please see the slide notes for details.

Figure 2. High-throughput sequencing for PS-THSP products of random sequences with a serial dilution. (A) PS-THSP was ... Figure 2. High-throughput sequencing for PS-THSP products of random sequences with a serial dilution. (A) PS-THSP was performed with a PS-THSP probe 1 (100 nM) and different amount of dNTPs (M: 50 bp DNA ladder, lane 1: 0 μM, lane 2: 2 μM, lane 3: 20 μM, lane 4: 200 μM, lane 5: 2 mM). The product of first round was 10 times serially diluted to fourth round and PS-THSP was performed for each round. (B) The flow chart shows that how the next generation sequencing data for PS-THSP products was analysed. (C) The four-way Venn diagram was prepared with the analysed NGS data for all the rounds. Unless provided in the caption above, the following copyright applies to the content of this slide: © The Author(s) 2019. Published by Oxford University Press on behalf of Nucleic Acids Research.This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com Nucleic Acids Res, Volume 47, Issue 5, 30 January 2019, Pages 2169–2176, https://doi.org/10.1093/nar/gkz044 The content of this slide may be subject to copyright: please see the slide notes for details.