Related Experiment Video
Updated: Feb 7, 2026
08:31
Rapid Assembly of Multi-Gene Constructs using Modular Golden Gate Cloning
Published on: February 5, 2021
15.0K
Slowpoke: An Automated Golden Gate Cloning Workflow for Opentrons OT-2 and Flex
Koray Malcı1,2, Fankang Meng1,2, Henri Galez3
1Department of Bioengineering, Imperial College London, London SW7 2AZ, U.K.
ACS Synthetic Biology
|February 5, 2026
Summary
Slowpoke automates Golden Gate cloning using open-source liquid handlers, simplifying DNA assembly for synthetic biology. This user-friendly workflow enhances standardization and high-throughput capacity in research labs.
Area of Science:
- Synthetic biology
- Molecular biology
- Bioengineering
Background:
- DNA assembly is crucial in synthetic biology, with growing needs for automation, standardization, and high-throughput methods.
- Existing solutions often lack accessibility or flexibility for diverse laboratory settings.
- The demand for error-free and efficient DNA assembly drives innovation in automated workflows.
Purpose of the Study:
- To present Slowpoke, a user-friendly and flexible automated workflow for Golden Gate DNA assembly.
- To adapt this workflow for popular, low-cost, open-source liquid-handling platforms (Opentrons OT-2 and Flex).
- To develop a graphical user interface (GUI) for simplified protocol generation.
Main Methods:
- Automated execution of DNA cloning, Escherichia coli transformation, plating, and colony PCR using Opentrons OT-2 and Flex.
- Development of a free, web-based GUI (https://slowpoke.streamlit.app/) for protocol creation via file uploads.
- Validation using the MoClo Yeast Toolkit (YTK) and SubtiToolKit (STK) for basic and high-throughput assemblies.
Main Results:
- High assembly efficiencies achieved: 17/17 (YTK/OT-2), 11/12 (YTK/Flex), and 8/13 (STK/OT-2) for basic constructs.
- Successful high-throughput assembly of six-part constructs using YTK-compatible parts on Flex (55/57 correct).
- Demonstrated robustness and adaptability across different toolkits and automation platforms.
Conclusions:
- Slowpoke provides an accessible, modular, and automated solution for Golden Gate cloning.
- The workflow significantly enhances standardization and high-throughput capacity in synthetic biology research.
- The freely available suite (https://github.com/Tom-Ellis-Lab/Slowpoke) supports broader adoption in research laboratories.
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