Automating the Illumina DNA library preparation kit for whole genome sequencing applications on the flowbot ONE liquid handler robot

Whole-genome sequencing (WGS) is currently making its transition from research tool into routine (clinical) diagnostic practice. The workflow for WGS includes the highly labor-intensive library preparations (LP), one of the most critical steps in the WGS procedure. Here, we describe the automation o...

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Bibliographic Details
Published in:Scientific reports 2024-04, Vol.14 (1), p.8159-8159, Article 8159
Main Authors: Meijers, Erin, Verhees, Fabienne B., Heemskerk, Dennis, Wessels, Els, Claas, Eric C. J., Boers, Stefan A.
Format: Article
Language:English
Subjects:
631/1647/514/2254
631/326
Automation
Deoxyribonucleic acid
DNA
Gene Library
Genomes
High-Throughput Nucleotide Sequencing - methods
Humanities and Social Sciences
Humans
Lipopolysaccharides
multidisciplinary
Multilocus sequence typing
Nucleotide sequence
Reproducibility
Reproducibility of Results
Risk reduction
Robotics
Robots
Science
Science (multidisciplinary)
Whole Genome Sequencing
Workflow
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Summary:Whole-genome sequencing (WGS) is currently making its transition from research tool into routine (clinical) diagnostic practice. The workflow for WGS includes the highly labor-intensive library preparations (LP), one of the most critical steps in the WGS procedure. Here, we describe the automation of the LP on the flowbot ONE robot to minimize the risk of human error and reduce hands-on time (HOT). For this, the robot was equipped, programmed, and optimized to perform the Illumina DNA Prep automatically. Results obtained from 16 LP that were performed both manually and automatically showed comparable library DNA yields (median of 1.5-fold difference), similar assembly quality values, and 100% concordance on the final core genome multilocus sequence typing results. In addition, reproducibility of results was confirmed by re-processing eight of the 16 LPs using the automated workflow. With the automated workflow, the HOT was reduced to 25 min compared to the 125 min needed when performing eight LPs using the manual workflow. The turn-around time was 170 and 200 min for the automated and manual workflow, respectively. In summary, the automated workflow on the flowbot ONE generates consistent results in terms of reliability and reproducibility, while significantly reducing HOT as compared to manual LP.
ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-024-58963-2