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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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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author Lorena Hägele
Brian F. Pfleger
Ralf Takors
author_facet Lorena Hägele
Brian F. Pfleger
Ralf Takors
author_sort Lorena Hägele
collection DOAJ
description 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.
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spelling doaj-art-780a77adf7f84ce9bd4d210c0f1a87e62025-08-20T01:56:00ZengMDPI AGBioengineering2306-53542024-09-0111989210.3390/bioengineering11090892Getting the Right Clones in an Automated Manner: An Alternative to Sophisticated Colony-Picking RoboticsLorena Hägele0Brian F. Pfleger1Ralf Takors2Institute of Biochemical Engineering, University of Stuttgart, 70569 Stuttgart, GermanyDepartment of Chemical and Biological Engineering, University of Wisconsin-Madison, Madison, WI 53706, USAInstitute of Biochemical Engineering, University of Stuttgart, 70569 Stuttgart, GermanyIn 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.https://www.mdpi.com/2306-5354/11/9/892laboratory automationstrain constructionhigh-throughput clone selectioncolony-picking stationbiofoundry
spellingShingle Lorena Hägele
Brian F. Pfleger
Ralf Takors
Getting the Right Clones in an Automated Manner: An Alternative to Sophisticated Colony-Picking Robotics
Bioengineering
laboratory automation
strain construction
high-throughput clone selection
colony-picking station
biofoundry
title Getting the Right Clones in an Automated Manner: An Alternative to Sophisticated Colony-Picking Robotics
title_full Getting the Right Clones in an Automated Manner: An Alternative to Sophisticated Colony-Picking Robotics
title_fullStr Getting the Right Clones in an Automated Manner: An Alternative to Sophisticated Colony-Picking Robotics
title_full_unstemmed Getting the Right Clones in an Automated Manner: An Alternative to Sophisticated Colony-Picking Robotics
title_short Getting the Right Clones in an Automated Manner: An Alternative to Sophisticated Colony-Picking Robotics
title_sort getting the right clones in an automated manner an alternative to sophisticated colony picking robotics
topic laboratory automation
strain construction
high-throughput clone selection
colony-picking station
biofoundry
url https://www.mdpi.com/2306-5354/11/9/892
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AT ralftakors gettingtherightclonesinanautomatedmanneranalternativetosophisticatedcolonypickingrobotics