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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...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2021
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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). |
format | Online Article Text |
id | pubmed-7994540 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT isogawaasako enrichmentofplasmidpoolfromecoliformasssequencingtodetectuntargetedmutagenicevents AT fuchsrobertp enrichmentofplasmidpoolfromecoliformasssequencingtodetectuntargetedmutagenicevents AT fujiishingo enrichmentofplasmidpoolfromecoliformasssequencingtodetectuntargetedmutagenicevents |