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Integrated Addressable Dynamic Droplet Array (aDDA) as Sub-Nanoliter Reactors for High-Coverage Genome Sequencing of Single Yeast Cells
Small ( IF 13.0 ) Pub Date : 2021-07-23 , DOI: 10.1002/smll.202100325 Chunyu Li 1 , Yanhai Gong 1 , Xixian Wang 1 , Jian Xu 1 , Bo Ma 1
Small ( IF 13.0 ) Pub Date : 2021-07-23 , DOI: 10.1002/smll.202100325 Chunyu Li 1 , Yanhai Gong 1 , Xixian Wang 1 , Jian Xu 1 , Bo Ma 1
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An addressable dynamic droplet array (aDDA) is presented that combines the advantages of static droplet arrays and continuous-flow droplet platforms. Modular fabrication is employed to create a self-contained integrated aDDA. All the sample preparation steps, including single-cell isolation, cell lysis, amplification, and product retrieval, are performed in sequence within a sub-nanoliter (≈300 pL) droplet. Sequencing-based validation suggests that aDDA reduces the amplification bias of multiple displacement amplification (MDA) and elevates the percentage of one-yeast-cell genome recovery to 91%, as compared to the average of 26% using conventional, 20 µL volume MDA reactions. Thus, aDDA is a valuable addition to the toolbox for high-genome-coverage sequencing of single microbial cells.
中文翻译：
集成可寻址动态液滴阵列 (aDDA) 作为亚纳升反应器，用于单酵母细胞的高覆盖基因组测序
提出了一种可寻址的动态液滴阵列 (aDDA)，它结合了静态液滴阵列和连续流动液滴平台的优点。采用模块化制造来创建独立的集成 aDDA。所有样品制备步骤，包括单细胞分离、细胞裂解、扩增和产物回收，均在亚纳升 (≈300 pL) 液滴内依次执行。基于测序的验证表明，与使用常规 20 µL 体积 MDA 反应的平均值 26% 相比，aDDA 降低了多重置换扩增 (MDA) 的扩增偏差，并将单酵母细胞基因组回收率提高到 91% . 因此，aDDA 是对单个微生物细胞的高基因组覆盖测序工具箱的宝贵补充。
更新日期：2021-09-16
中文翻译：
集成可寻址动态液滴阵列 (aDDA) 作为亚纳升反应器，用于单酵母细胞的高覆盖基因组测序
提出了一种可寻址的动态液滴阵列 (aDDA)，它结合了静态液滴阵列和连续流动液滴平台的优点。采用模块化制造来创建独立的集成 aDDA。所有样品制备步骤，包括单细胞分离、细胞裂解、扩增和产物回收，均在亚纳升 (≈300 pL) 液滴内依次执行。基于测序的验证表明，与使用常规 20 µL 体积 MDA 反应的平均值 26% 相比，aDDA 降低了多重置换扩增 (MDA) 的扩增偏差，并将单酵母细胞基因组回收率提高到 91% . 因此，aDDA 是对单个微生物细胞的高基因组覆盖测序工具箱的宝贵补充。
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