AI Article Synopsis
- Golden Gate cloning is a widely-used DNA assembly method known for its modular design, enabling the creation of complex DNA fragments and a repository of reusable components, which minimizes validation costs.
- Increasing the number of reactions may raise consumable costs and human error potential, as typical reactions are done in volumes of 10-25 μL.
- Recent advances in liquid handling technology, specifically acoustic dispensers, allow for miniaturized reactions in a sustainable 1 μL volume, reducing plastic waste and reagent usage while optimizing the cloning process.
Article Abstract
Golden Gate cloning has become one of the most popular DNA assembly techniques. Its modular and hierarchical structure allows the construction of complex DNA fragments. Over time, Golden Gate cloning allows for the creation of a repository of reusable parts, reducing the cost of frequent sequence validation. However, as the number of reactions and fragments increases, so does the cost of consumables and the potential for human error. Typically, Golden Gate reactions are performed in volumes of 10-25 μL. Recent technological advances have led to the development of liquid handling robots that use sound to transfer liquids in the nL range from a source plate to a target plate. These acoustic dispensers have become particularly popular in the field of synthetic biology. The use of this technology allows miniaturization and parallelization of molecular reactions in a tip-free manner, making it sustainable by reducing plastic waste and reagent usage. Here, we provide a step-by-step protocol for performing and parallelizing Golden Gate cloning reactions in 1 μL total volume.
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| http://dx.doi.org/10.1007/978-1-0716-4220-7_9 | DOI Listing |
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