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

富集大肠杆菌质粒池进行大规模测序,以检测非靶向诱变事件

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作者:Asako Isogawa, Robert P Fuchs, Shingo Fujii

Abstract

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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