TeselaGen Biotechnology, Inc., San Francisco, CA, USA.
DOE Joint Genome Institute (JGI), Walnut Creek, CA, USA.
Methods Mol Biol. 2020;2205:19-47. doi: 10.1007/978-1-0716-0908-8_2.
Modern DNA assembly techniques are known for their potential to link multiple large DNA fragments together into even larger constructs in single pot reactions that are easier to automate and work more reliably than traditional cloning methods. The simplicity of the chemistry is in contrast to the increased work needed to design optimal reactions that maximize DNA fragment reuse, minimize cost, and organize thousands of potential chemical reactions. Here we examine available DNA assembly methods and describe through example, the construction of a complex but not atypical combinatorial and hierarchical library using protocols that are generated automatically with the assistance of modern synthetic biology software.
现代 DNA 组装技术以其将多个大型 DNA 片段在单管反应中连接成更大构建体的潜力而闻名，这种反应比传统的克隆方法更容易自动化，并且更可靠。与增加的工作量形成对比的是，化学的简单性在于设计最佳反应以最大化 DNA 片段的再利用、最小化成本和组织数千种潜在化学反应的需要。在这里，我们检查了可用的 DNA 组装方法，并通过示例描述了使用在现代合成生物学软件的帮助下自动生成的方案构建复杂但非典型的组合和层次文库的过程。
