CNRS, UMR7258, Marseille 13009, France.
Inserm, U1068, Marseille 13009, France.
STAR Protoc. 2021 Mar 20;2(2):100399. doi: 10.1016/j.xpro.2021.100399. eCollection 2021 Jun 18.
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 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 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).
跨损伤合成(TLS)是一种应对 DNA 损伤的事件。在 TLS 过程中,负责的 TLS 聚合酶经常引发非靶向性突变,这可能是遗传多样性的来源。由于大部分 DNA 不经历 TLS,因此鉴定这种非靶向性突变具有挑战性。在这里,我们提出了一种在 中富集经历 Pol V 介导的 TLS 的质粒池的方案,以便进行大规模测序。该方案的概念可应用于任何物种。有关此方案使用和执行的完整详细信息,请参阅 Isogawa 等人。(2018)。