Protein engineering and synthetic biology applications increasingly rely on the assembly of modular libraries composed of thousands of different combinations of DNA building blocks. At present, the validation of such libraries is performed by Sanger sequencing analysis on a small subset of clones on an ad hoc basis. Here, we implement a systematic procedure for the comprehensive evaluation of combinatorial libraries, immediately after their creation in vitro, using long reads sequencing technology. After an initial step of nanopore sequencing, we use straightforward bioinformatics tools to tabulate the composition and synteny of the building blocks in each read. We subsequently use exploratory statistics to assess the library and validate its diversity before carrying downstream cloning and screening assays.

中文翻译：

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使用纳米孔测序对组合 DNA 文库进行质量控制和统计评估。

蛋白质工程和合成生物学应用越来越依赖于由数千种不同的 DNA 构建块组合组成的模块化文库的组装。目前，此类文库的验证是通过 Sanger 测序分析临时对一小部分克隆进行的。在这里，我们在体外创建组合文库后立即实施系统程序，对组合文库进行综合评估，使用长读长测序技术。在纳米孔测序的初始步骤之后，我们使用简单的生物信息学工具将每次读取中构建块的组成和同线性制成表格。我们随后使用探索性统计来评估文库并在进行下游克隆和筛选分析之前验证其多样性。