Peter Vegh, Elliott Chapman, Craig Gilmour, Rennos Fragkoudis
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引用次数: 0
摘要
金门克隆技术能将 DNA 部分模块化地组装成所需的合成基因构建体。金门反应的 "一锅 "性质使其特别适合于高通量自动化,便于以大规模并行的方式生成成千上万的构建体。这一过程中的一个潜在瓶颈是这些构建体的设计。在设计装配流程时必须考虑多个参数,而且在实施前还应检查和验证最终设计。对大量结构体进行手工操作既不实际也不可行,而且会增加引入潜在代价高昂的错误的可能性。在本章中,我们将介绍一种设计工作流程,它利用定制的计算工具自动完成构建体设计流程的关键阶段,并分批进行序列编辑。
Modular DNA Construct Design for High-Throughput Golden Gate Assembly.
Golden Gate cloning enables the modular assembly of DNA parts into desired synthetic genetic constructs. The "one-pot" nature of Golden Gate reactions makes them particularly amenable to high-throughput automation, facilitating the generation of thousands of constructs in a massively parallel manner. One potential bottleneck in this process is the design of these constructs. There are multiple parameters that must be considered during the design of an assembly process, and the final design should also be checked and verified before implementation. Doing this by hand for large numbers of constructs is neither practical nor feasible and increases the likelihood of introducing potentially costly errors. In this chapter we describe a design workflow that utilizes bespoke computational tools to automate the key phases of the construct design process and perform sequence editing in batches.
期刊介绍:
For over 20 years, biological scientists have come to rely on the research protocols and methodologies in the critically acclaimed Methods in Molecular Biology series. The series was the first to introduce the step-by-step protocols approach that has become the standard in all biomedical protocol publishing. Each protocol is provided in readily-reproducible step-by-step fashion, opening with an introductory overview, a list of the materials and reagents needed to complete the experiment, and followed by a detailed procedure that is supported with a helpful notes section offering tips and tricks of the trade as well as troubleshooting advice.