Low-input PacBio sequencing generates high-quality individual fly genomes and characterizes mutational processes

文献类型： 外文期刊

第一作者： Jia, Hangxing

作者： Jia, Hangxing;Tan, Shengjun;Cai, Yingao;Guo, Yanyan;Shen, Jieyu;Zhang, Yaqiong;Ma, Huijing;Zhang, Qingzhu;Qiao, Gexia;Zhang, Yong E.;Cai, Yingao;Guo, Yanyan;Shen, Jieyu;Zhang, Qingzhu;Chen, Jinfeng;Qiao, Gexia;Zhang, Yong E.;Chen, Jinfeng;Ruan, Jue

作者机构：

期刊名称：NATURE COMMUNICATIONS （ 影响因子：14.7； 五年影响因子：16.1 ）

ISSN：

年卷期： 2024 年 15 卷 1 期

页码：

收录情况： SCI

摘要： Long-read sequencing, exemplified by PacBio, revolutionizes genomics, overcoming challenges like repetitive sequences. However, the high DNA requirement ( > 1 mu g) is prohibitive for small organisms. We develop a low-input (100 ng), low-cost, and amplification-free library-generation method for PacBio sequencing (LILAP) using Tn5-based tagmentation and DNA circularization within one tube. We test LILAP with two Drosophila melanogaster individuals, and generate near-complete genomes, surpassing preexisting single-fly genomes. By analyzing variations in these two genomes, we characterize mutational processes: complex transpositions (transposon insertions together with extra duplications and/or deletions) prefer regions characterized by non-B DNA structures, and gene conversion of transposons occurs on both DNA and RNA levels. Concurrently, we generate two complete assemblies for the endosymbiotic bacterium Wolbachia in these flies and similarly detect transposon conversion. Thus, LILAP promises a broad PacBio sequencing adoption for not only mutational studies of flies and their symbionts but also explorations of other small organisms or precious samples.

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