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

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

Carla Teixeira  1   2   3 , Andrey Ipatov  1 , Joana Carvalho  1   4 , Agnes Purwidyantri  1   5 , Natacha Fontes  6 , Marta Prado  1
Affiliations

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

Carla Teixeira et al. ACS Omega. .

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%.

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Conflict of interest statement

The authors declare no competing financial interest.

Figures

Figure 1
Figure 1
Miniaturized DNA-purification device module. (A) Prototype set: on the left, PMDS disposable cartridge and glass microfiber filter in the center; on the right, PMMA support. (B) Open view of the prototype. (C) Top view of the assembled prototype. (D) Diagonal view prototype with the fluid inlet and outlet channels.
Figure 2
Figure 2
Protocols (P1, P2, P2.1, and P3). λDNA total mass, ng, was collected after each extraction step. Results are represented as mean ± 90% CI, n = 3, independent replicates. Results show a high DNA loss in binding for P1 and a high DNA recovery in elution for the remaining protocols.
Figure 3
Figure 3
DNA extraction and purification from leaves and grapes with the DNeasy PowerSoil Pro Kit (Qiagen). Recovered DNA concentration, in ng μL–1, for each grapevine variety is represented. Results are shown as mean ± 90% CI, n = 9, independent and dependent replicates. Results show a significant difference in the concentration of DNA extracted between leaves and grapes in most varieties.
Figure 4
Figure 4
DNA extraction and purification from leaves with the miniaturized device and P2.1. Graphic shows recovered DNA concentration, ng μL–1, for each variety and evolution of DNA recovery along the elution (E1–E5). Results are represented as mean ± 90% CI, n = 9, independent and dependent replicates.
Figure 5
Figure 5
DNA extraction and purification from leaves with the miniaturized device and P2.1. Graphic shows recovered DNA concentration, ng μL–1, for each variety and the evolution of DNA recovery along the elution (E1–E5). Results are represented as mean ± 90% CI, n = 9, independent and dependent replicates.
See this image and copyright information in PMC

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