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. 2020 Aug 19;10:13981. doi: 10.1038/s41598-020-70841-1

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© The Author(s) 2020

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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Schematic of the CAVE pipeline for bacteriophage directed evolution, with iterative cycles of mutagenesis and thermal selection. In this procedure, bases within the bacteriophage genomes are randomly affected by treatment with a chemical mutagen. Upon replication within a host, a mismatch mutation is introduced. The mutant library is then subjected to high-temperature incubation to select for thermally stable variants; surviving phages are amplified and carried over to subsequent iterations.