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Enrichment of plasmid pool from E. coli for mass sequencing to detect untargeted mutagenic events

Translesion synthesis (TLS) is an event to cope with DNA damages. During TLS, the responsible TLS polymerase frequently elicits untargeted mutagenesis as potentially a source of genetic diversity. Identifying such untargeted mutations in vivo is challenging due to the bulk of DNA that does not under...

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Detalles Bibliográficos
Autores principales: Isogawa, Asako, Fuchs, Robert P., Fujii, Shingo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7994540/
https://www.ncbi.nlm.nih.gov/pubmed/33786464
http://dx.doi.org/10.1016/j.xpro.2021.100399
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author Isogawa, Asako
Fuchs, Robert P.
Fujii, Shingo
author_facet Isogawa, Asako
Fuchs, Robert P.
Fujii, Shingo
author_sort Isogawa, Asako
collection PubMed
description Translesion synthesis (TLS) is an event to cope with DNA damages. During TLS, the responsible TLS polymerase frequently elicits untargeted mutagenesis as potentially a source of genetic diversity. Identifying such untargeted mutations in vivo is challenging due to the bulk of DNA that does not undergo TLS. Here, we present a protocol to enrich a plasmid pool that underwent Pol V-mediated TLS in Escherichia coli for mass sequencing. The concept of this protocol could be applied into any species. For complete details on the use and execution of this protocol, please refer to Isogawa et al. (2018).
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spelling pubmed-79945402021-03-29 Enrichment of plasmid pool from E. coli for mass sequencing to detect untargeted mutagenic events Isogawa, Asako Fuchs, Robert P. Fujii, Shingo STAR Protoc Protocol Translesion synthesis (TLS) is an event to cope with DNA damages. During TLS, the responsible TLS polymerase frequently elicits untargeted mutagenesis as potentially a source of genetic diversity. Identifying such untargeted mutations in vivo is challenging due to the bulk of DNA that does not undergo TLS. Here, we present a protocol to enrich a plasmid pool that underwent Pol V-mediated TLS in Escherichia coli for mass sequencing. The concept of this protocol could be applied into any species. For complete details on the use and execution of this protocol, please refer to Isogawa et al. (2018). Elsevier 2021-03-20 /pmc/articles/PMC7994540/ /pubmed/33786464 http://dx.doi.org/10.1016/j.xpro.2021.100399 Text en © 2021 The Author(s) http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Isogawa, Asako
Fuchs, Robert P.
Fujii, Shingo
Enrichment of plasmid pool from E. coli for mass sequencing to detect untargeted mutagenic events
title Enrichment of plasmid pool from E. coli for mass sequencing to detect untargeted mutagenic events
title_full Enrichment of plasmid pool from E. coli for mass sequencing to detect untargeted mutagenic events
title_fullStr Enrichment of plasmid pool from E. coli for mass sequencing to detect untargeted mutagenic events
title_full_unstemmed Enrichment of plasmid pool from E. coli for mass sequencing to detect untargeted mutagenic events
title_short Enrichment of plasmid pool from E. coli for mass sequencing to detect untargeted mutagenic events
title_sort enrichment of plasmid pool from e. coli for mass sequencing to detect untargeted mutagenic events
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7994540/
https://www.ncbi.nlm.nih.gov/pubmed/33786464
http://dx.doi.org/10.1016/j.xpro.2021.100399
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