Getting the Right Clones in an Automated Manner: An Alternative to Sophisticated Colony-Picking Robotics

In recent years, the design–build–test–learn (DBTL) cycle has become a key concept in strain engineering. Modern biofoundries enable automated DBTL cycling using robotic devices. However, both highly automated facilities and semi-automated facilities encounter bottlenecks in clone selection and scre...

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Bibliographic Details
Main Authors: Lorena Hägele, Brian F. Pfleger, Ralf Takors
Format: Article
Language:English
Published: MDPI AG 2024-09-01
Series:Bioengineering
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Online Access:https://www.mdpi.com/2306-5354/11/9/892
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Summary:In recent years, the design–build–test–learn (DBTL) cycle has become a key concept in strain engineering. Modern biofoundries enable automated DBTL cycling using robotic devices. However, both highly automated facilities and semi-automated facilities encounter bottlenecks in clone selection and screening. While fully automated biofoundries can take advantage of expensive commercially available colony pickers, semi-automated facilities have to fall back on affordable alternatives. Therefore, our clone selection method is particularly well-suited for academic settings, requiring only the basic infrastructure of a biofoundry. The automated liquid clone selection (ALCS) method represents a straightforward approach for clone selection. Similar to sophisticated colony-picking robots, the ALCS approach aims to achieve high selectivity. Investigating the time analogue of five generations, the model-based set-up reached a selectivity of 98 ± 0.2% for correctly transformed cells. Moreover, the method is robust to variations in cell numbers at the start of ALCS. Beside <i>Escherichia coli</i>, promising chassis organisms, such as <i>Pseudomonas putida</i> and <i>Corynebacterium glutamicum</i>, were successfully applied. In all cases, ALCS enables the immediate use of the selected strains in follow-up applications. In essence, our ALCS approach provides a ‘low-tech’ method to be implemented in biofoundry settings without requiring additional devices.
ISSN:2306-5354