利用液体处理仪在海洋模式无脊椎动物中进行高通量基因表达谱分析。

IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY BMC Biotechnology Pub Date : 2024-01-19 DOI:10.1186/s12896-024-00831-y
Giovanni Annona, Assunta Liberti, Carla Pollastro, Antonietta Spagnuolo, Paolo Sordino, Pasquale De Luca
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引用次数: 0

摘要

背景:现代高通量技术可同时处理大量样本,并提供快速准确的程序。近年来,自动液体处理工作站已成为可重复样品制备的成熟技术。它们具有灵活性,适用于越来越多的应用领域。通常,这种方法主要用于细胞系处理和异种生物测试的实验程序。相反,人们很少关注自动液体处理仪在整个生物体分析中的应用,这通常涉及耗时的实验室程序:结果:在此,我们介绍了一种全自动工作流程,从 RNA 提取到实时 PCR 处理,所有步骤都是自动化的,用于定量检测海洋生物模型 Ciona robusta 的基因表达。在程序验证方面,我们比较了液体处理机与传统手工程序的结果。结果显示,两者的结果相当,尤其是在样品处理的初始步骤中节省了大量时间:结论:这项工作将该技术的可能应用领域扩展到全身生物体,减轻了手工操作可能产生的问题。通过减少错误、避免交叉污染、缩短操作时间和简化程序,该技术可用于大规模筛选调查。
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Reaping the benefits of liquid handlers for high-throughput gene expression profiling in a marine model invertebrate.

Background: 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.

Results: 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.

Conclusions: 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.

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来源期刊
BMC Biotechnology
BMC Biotechnology 工程技术-生物工程与应用微生物
CiteScore
6.60
自引率
0.00%
发文量
34
审稿时长
2 months
期刊介绍: BMC Biotechnology is an open access, peer-reviewed journal that considers articles on the manipulation of biological macromolecules or organisms for use in experimental procedures, cellular and tissue engineering or in the pharmaceutical, agricultural biotechnology and allied industries.
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