Department of Biological Systems Engineering, Virginia Polytechnic Institute and State University, Blacksburg, VA, USA.
Methods Mol Biol. 2025;2850:133-147. doi: 10.1007/978-1-0716-4220-7_8.
Golden Gate assembly is a requisite method in synthetic biology that facilitates critical conventions such as genetic part abstraction and rapid prototyping. However, compared to robotic implementation, manual Golden Gate implementation is cumbersome, error-prone, and inconsistent for complex assembly designs. AssemblyTron is an open-source python package that provides an affordable automation solution using open-source OpenTrons OT-2 lab robots. Automating Golden Gate assembly with AssemblyTron can reduce failure-rate, resource consumption, and training requirements for building complex DNA constructs, as well as indexed and combinatorial libraries. Here, we dissect a panel of upgrades to AssemblyTron's Golden Gate assembly capabilities, which include Golden Gate assembly into modular cloning part vectors, error-prone polymerase chain reaction (PCR) combinatorial mutant library assembly, and modular cloning indexed plasmid library assembly. These upgrades enable a broad pool of users with varying levels of experience to readily implement advanced Golden Gate applications using low-cost, open-source lab robotics.
金门组装是合成生物学中一种必要的方法，它促进了遗传部件抽象和快速原型制作等关键惯例。然而，与机器人实现相比，手动金门实现繁琐、容易出错，并且不适合复杂的组装设计。AssemblyTron 是一个开源的 Python 包，它使用开源的 OpenTrons OT-2 实验室机器人提供了一种负担得起的自动化解决方案。使用 AssemblyTron 自动化金门组装可以降低构建复杂 DNA 构建体、索引和组合文库的故障率、资源消耗和培训要求。在这里，我们剖析了 AssemblyTron 的金门组装功能的一系列升级，包括将金门组装到模块化克隆部件载体中、易错聚合酶链反应 (PCR) 组合突变文库组装以及模块化克隆索引质粒文库组装。这些升级使具有不同经验水平的广泛用户能够使用低成本、开源实验室机器人轻松实现高级金门应用。
