We present a technically simple, easy-to-perform method for generating the genomic libraries for Himar-1 transposon site sequencing (Tn-seq). In addition to being simpler than present methods in the technical aspect, it also allows more robust and straightforward identification of the insertion site, by generating a longer sequence surrounding the insertion TA in the genome. The method makes Tn-seq more user-friendly and accessible to laboratories with more-limited bioinformatic resources. Finally, we created a saturated transposon-mutant library in Mycobacterium abscessus and demonstrated the usefulness of the method in analysis of genes involved in colony morphology, as well as in analysis of the whole Tn-mutant library, with identification of over 8,000 unique mutants.

中文翻译：

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细菌中 Himar-1 转座子测序的一种简化且有效的方法，通过创建和分析脓肿分枝杆菌中的饱和转座子突变体库来证明

我们提出了一种技术简单、易于执行的方法，用于为 Himar-1 转座子位点测序 (Tn-seq) 生成基因组文库。除了在技术方面比现有方法更简单外，它还可以通过在基因组中围绕插入 TA 生成更长的序列来更可靠和直接地识别插入位点。该方法使 Tn-seq 对用户更加友好，并且可供生物信息资源有限的实验室使用。最后，我们在脓肿分枝杆菌中创建了一个饱和的转座子突变体库，并证明了该方法在分析涉及菌落形态的基因以及整个 Tn 突变体库的分析中的有用性，并鉴定了 8,000 多个独特的突变体。