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基于微型微尺度固相萃取的模块,用于从葡萄样品中高效提取和纯化DNA。

Miniaturized Microscale Solid-Phase Extraction-Based Module for Highly Efficient DNA Extraction and Purification from Grapevine Samples.

作者信息

Teixeira Carla, Ipatov Andrey, Carvalho Joana, Purwidyantri Agnes, Fontes Natacha, Prado Marta

机构信息

International Iberian Nanotechnology Laboratory (INL), Avenida Mestre José Veiga, 4715-310 Braga, Portugal.

Department of Biological Engineering, University of Minho, Campus de Gualtar, 4710-057 Braga, Portugal.

出版信息

ACS Omega. 2023 Aug 24;8(35):31738-31746. doi: 10.1021/acsomega.3c02717. eCollection 2023 Sep 5.

Abstract

Several developments over the last few years are being directed toward improving DNA-based analysis to simplify, miniaturize, and reduce the time and cost of analysis, with the objective to allow its use in decentralized settings. One of the most interesting fields is DNA extraction and purification, a key step for ensuring good analytical performance. In this sense, microscale solid phase extraction (μSPE) offers paramount advantages for an improved DNA yield. In this work, we have developed a miniaturized module for DNA purification based on μSPE using a borosilicate glass microfiber filter as the solid phase. We also established a protocol for highly efficient DNA purification from vegetable samples, including leaves and grapes from four different varieties from the PDO Douro and two varieties from the Minho wine regions. The protocol demonstrated excellent performance when compared with a commercial kit with a DNA recovery yield of around 50%.

摘要

过去几年的多项进展都致力于改进基于DNA的分析,以实现分析过程的简化、小型化,并减少分析时间和成本,目标是使其能够在分散的环境中使用。最有趣的领域之一是DNA提取和纯化,这是确保良好分析性能的关键步骤。从这个意义上说,微尺度固相萃取(μSPE)在提高DNA产量方面具有至关重要的优势。在这项工作中,我们开发了一种基于μSPE的小型化DNA纯化模块,使用硼硅酸盐玻璃微纤维过滤器作为固相。我们还建立了一种从蔬菜样品(包括来自杜罗法定产区四个不同品种的叶子和葡萄以及来自米尼奥葡萄酒产区的两个品种)中高效纯化DNA的方案。与商业试剂盒相比,该方案表现出色,DNA回收率约为50%。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f386/10483517/ee80876eb3a4/ao3c02717_0001.jpg

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