Journal
TALANTA
Volume 226, Issue -, Pages -Publisher
ELSEVIER
DOI: 10.1016/j.talanta.2021.122109
Keywords
Microfluidic device; DNA purification; Invasive species; eDNA; Dreissena polymorpha
Categories
Funding
- Confederacion Hidrografica del Guadalquivir
- INL
- Norte Portugal Regional Operational Programme (NORTE2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund [NORTE-01-0145-FEDER-000019]
- European Regional Development Fund (ERDP) under the EP -INTERREG V A Espana Portugal (POCTEP) [0181_NANOEATERS_1_e]
- FCT Fundacao para Ciencia e Tecnologia [SFRH/BD/140396/2018]
- Fundação para a Ciência e a Tecnologia [SFRH/BD/140396/2018] Funding Source: FCT
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Purification and concentration of DNA is crucial for DNA-based analysis. The novel miniaturized disposable device showed high efficiency in DNA purification, including handling highly diluted samples and environmental DNA with great potential.
Purification and concentration of DNA is a critical step on DNA-based analysis, which should ensure efficient DNA isolation and effective removal of contaminants that may interfere with downstream DNA amplification. Complexity of samples, minute content of target analyte, or high DNA fragmentation greatly entangles the success of this step. To overcome this issue, we designed and fabricated a novel miniaturized disposable device for a highly efficient DNA purification. The microfluidic device showed binding efficiency and elution yield of 90.1% and 86.7%, respectively. Moreover, the effect of DNA fragmentation, a parameter that has not been previously addressed, showed a great impact in the recovery step. The microfluidic system integrated micropillars with chitosan being used as the solid-phase for a pH-dependent DNA capture and release. We have showed the potential of the device in the successful purification of environmental DNA (eDNA) from river water samples contaminated with Dreissena polymorpha, an invasive alien species responsible for unquestionable economic and environmental consequences in river water basins. Additionally, the device was also able to concentrate the DNA extract from highly diluted samples, showing promising results for the early detection of such invasive species, which may allow prompt measures for a more efficient control in affected areas. Suitability for integration with downstream DNA analysis was also demonstrated through qPCR analysis of the samples purified with the microfluidic device, allowing detection of the target species even if highly diluted.
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