Novel approach for accurate minute DNA quantification on microvolumetric solutions

Carvalho, Joana; Negrinho, Renato; Azinheiro, Sarah; Garrido‐Maestu, Alejandro; Barros‐Velázquez, Jorge; Prado, Marta

doi:10.1016/j.microc.2018.02.001

Cited by 9 publications

(7 citation statements)

References 30 publications

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“…In P1, the initial DNA mass was obtained from quantification of the BB1 + λDNA (1:1) solution used for binding. For the remaining protocols (P2, P2.1, and P3), the initial DNA amount was estimated from the P1 results since the binding solution used in these protocolsBB2contains chaotropic salts that have previously shown to interfere with DNA quantification . Results are presented as mean ± 90% confidence interval (CI), n = 3 independent replicates.…”

Section: Methodsmentioning

confidence: 99%

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

Teixeira,

Ipatov,

Carvalho

et al. 2023

ACS Omega

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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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Section: Methodsmentioning

confidence: 99%

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

Teixeira,

Ipatov,

Carvalho

et al. 2023

ACS Omega

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“…The standard curves required for DNA quantification were obtained using NanoDrop 3300 software and compared for both DNA types. Regarding the salmon sperm DNA, the three mathematical models described in the related research article [1] (least squares, weighted least squares and weighted ridge regressions) were implemented, being these curves compared with the one obtained using NanoDrop 3300 software. In order to evaluate the precision of these mathematical models under varied conditions, in terms of repeatability, a total of 10 assays were performed and the relative standard deviation (% RSD) was calculated as an indication of precision regarding variations from assay to assay.…”

Section: Experimental Design Materials and Methodsmentioning

confidence: 99%

Data on minute DNA quantification on microvolumetric solutions: comparison of mathematical models and effect of some compounds on the DNA quantification accuracy

et al. 2018

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This article contains data related to the research article entitled “Novel approach for accurate minute DNA quantification on microvolumetric solutions” (Carvalho et al., 2018). The combination of PicoGreen® with a microvolume fluorospectrometer is a popular DNA quantification method due to its high sensitivity and minimal consumption of sample, being commonly used to evaluate the performance of microfluidic devices designed for DNA purification. In this study, the authors present data related with the effect of DNA fragmentation level. The present data article includes the data used on the precision evaluation, in terms of repeatability, of the mathematical models developed to obtain the standards curve for salmon sperm DNA (low molecular weight). In addition, results related with the effect of some compounds on the DNA quantification accuracy using λDNA are presented.

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“…In this sense, the use of microfluidic devices and nanoparticles is already improving DNA recovery and extraction processes from complex food matrices. 112,113 Besides advances in technologies and instrumentation, issues of method development, validation, and harmonization have to be addressed in terms of assuring the validity of future studies prior to the implementation of the developed methods in the industry: (i) all of the possible sources of variability, such as changes caused by intraspecies differences or food processing, need to be controlled; (ii) power analysis should be performed to check statistical validity of the results; (iii) interlaboratory comparisons should be addressed in order to guarantee that comparable analytical results are provided; and (iv) certified reference materials and operation procedures should be developed and made available for method standardization.…”

Section: ■ Lampmentioning

confidence: 99%

“…Because food samples frequently contain fragmented DNA and minute concentrations, new sample-preparation protocols able to deal with these trace-level DNA amounts will also be needed. In this sense, the use of microfluidic devices and nanoparticles is already improving DNA recovery and extraction processes from complex food matrices. , …”

Section: Concluding Remarks and Future Trendsmentioning

confidence: 99%

Review of Recent DNA-Based Methods for Main Food-Authentication Topics

Böhme

Calo-Mata

Barros‐Velázquez

et al. 2019

J. Agric. Food Chem.

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156

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Adulteration and mislabeling of food products and the commercial fraud derived, either intentionally or not, is a global source of economic fraud to consumers but also to all stakeholders involved in food production and distribution. Legislation has been enforced all over the world aimed at guaranteeing the authenticity of the food products all along the distribution chain, thereby avoiding food fraud and adulteration. Accordingly, there is a growing need for new analytical methods able to verify that all the ingredients included in a foodstuff match the qualities claimed by the manufacturer or distributor. In this sense, the improved performance of most recent DNA-based tools in term of sensitivity, multiplexing ability, high-throughput, and relatively low-cost give them a game-changing role in food-authenticity-related topics. Here, we provide a thorough and updated vision on the recently reported approaches that are applying these DNA-based tools to assess the authenticity of food components and products.

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Novel approach for accurate minute DNA quantification on microvolumetric solutions

Cited by 9 publications

References 30 publications

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

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

Data on minute DNA quantification on microvolumetric solutions: comparison of mathematical models and effect of some compounds on the DNA quantification accuracy

Review of Recent DNA-Based Methods for Main Food-Authentication Topics

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