DNA-BOT: A low-cost, automated DNA assembly platform for synthetic biology
DNA-BOT: A low-cost, automated DNA assembly platform for synthetic biology
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DNA-BOT：用于合成生物学的低成本、自动化 DNA 组装平台
DOI：
10.1101/832139
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发表时间：
2019
期刊：
影响因子：
--
通讯作者：
Storch M
中科院分区：
文献类型：
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作者：
Storch M
多部分 DNA 组装是合成和分子生物学中许多项目的物理起点。通过构建广泛的 DNA 构建体库来探索遗传设计空间的能力对于创建编程生物系统至关重要。随着合成生物学界采用多种 DNA 组装方法和标准，DNA 组装过程的自动化现已受到广泛关注。自动化将使用更少的研究人员时间实现更大的构建，同时增加可访问的设计空间。然而，这些好处目前导致设备和消耗品成本高昂。在这里，我们通过在 OpenTrons (DNA-BOT) 上使用 BASIC 引入低成本 DNA 组装来解决这一限制。为此，我们开发了一个开源软件包，并通过同时组装由10个遗传部分组成的88个构建体，评估三基因操纵子的启动子、核糖体结合位点和基因顺序设计空间，展示了DNA-BOT的性能。所有 88 个结构均以高精度组装，每个结构的耗材成本为 1.50 至 5.50 美元。这说明了 DNA-BOT 的效率、准确性和经济性，使其可供大多数实验室使用，并使自动化 DNA 组装大众化。
Multi-part DNA assembly is the physical starting point for many projects in Synthetic and Molecular Biology. The ability to explore a genetic design space by building extensive libraries of DNA constructs is essential for creating programmed biological systems. With multiple DNA assembly methods and standards adopted in the Synthetic Biology community, automation of the DNA assembly process is now receiving serious attention. Automation will enable larger builds using less researcher time, while increasing the accessible design space. However, these benefits currently incur high costs for both equipment and consumables. Here, we address this limitation by introducing low-cost DNA assembly with BASIC on OpenTrons (DNA-BOT). For this purpose, we developed an open-source software package and demonstrated the performance of DNA-BOT by simultaneously assembling 88 constructs composed of 10 genetic parts, evaluating the promoter, ribosome binding site and gene order design space for a three-gene operon. All 88 constructs were assembled with high accuracy, at a consumables cost of $1.50–$5.50 per construct. This illustrates the efficiency, accuracy and affordability of DNA-BOT, making it accessible for most labs and democratizing automated DNA assembly.