Reaping the benefits of liquid handlers for high-throughput gene expression profiling in a marine model invertebrate

• Published in: BMC biotechnology Vol. 24; no. 1; p. 4
• Main Authors: Annona, Giovanni, Liberti, Assunta, Pollastro, Carla, Spagnuolo, Antonietta, Sordino, Paolo, De Luca, Pasquale
• Format: Journal Article
• Language: English
• Published: England BioMed Central Ltd 19.01.2024; BioMed Central; BMC
• Subjects: Analysis; Automated protocols; Automation; Biological models; Biological research; Biology, Experimental; Ciona robusta; Contamination; Extraction (Chemistry); Gene expression; Gene Expression Profiling; Genes; Genetic aspects; Genomes; Handlers; High-throughput screening (Biochemical assaying); Immune system; Instrument industry; Laboratory automation; Large-scale screening; Marine invertebrates; Marine organisms; Marine technology; Methods; Microarray Analysis; Minors; Molecular biology; Organisms; Polymerase chain reaction; Protocol; Real-Time Polymerase Chain Reaction; RNA; Robotics; Sample preparation; Specimen Handling - methods; Technology application; Workflow; Workstations; Xenobiotics; United States; Italy; Large-scale screening; Ciona robusta; Laboratory automation; Automated protocols; Marine organisms; Molecular biology protocols; Immune system
• ISSN: 1472-6750; 1472-6750
• DOI: 10.1186/s12896-024-00831-y

Modern high-throughput technologies enable the processing of a large number of samples simultaneously, while also providing rapid and accurate procedures. In recent years, automated liquid handling workstations have emerged as an established technology for reproducible sample preparation. They offer flexibility, making them suitable for an expanding range of applications. Commonly, such approaches are well-developed for experimental procedures primarily designed for cell-line processing and xenobiotics testing. Conversely, little attention is focused on the application of automated liquid handlers in the analysis of whole organisms, which often involves time-consuming laboratory procedures. Here, we present a fully automated workflow for all steps, from RNA extraction to real-time PCR processing, for gene expression quantification in the ascidian marine model Ciona robusta. For procedure validation, we compared the results obtained with the liquid handler with those of the classical manual procedure. The outcome revealed comparable results, demonstrating a remarkable time saving particularly in the initial steps of sample processing. This work expands the possible application fields of this technology to whole-body organisms, mitigating issues that can arise from manual procedures. By minimizing errors, avoiding cross-contamination, decreasing hands-on time and streamlining the procedure, it could be employed for large-scale screening investigations.