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Magnetophoresis in Centrifugal Microfluidics at Continuous Rotation for Nucleic Acid Extraction

Centrifugal microfluidics enables fully automated molecular diagnostics at the point-of-need. However, the integration of solid-phase nucleic acid extraction remains a challenge. Under this scope, we developed the magnetophoresis under continuous rotation for magnetic bead-based nucleic acid extract...

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Detalles Bibliográficos
Autores principales: Hin, Sebastian, Paust, Nils, Rombach, Markus, Lüddecke, Jan, Specht, Mara, Zengerle, Roland, Mitsakakis, Konstantinos
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9787563/
https://www.ncbi.nlm.nih.gov/pubmed/36557411
http://dx.doi.org/10.3390/mi13122112
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author Hin, Sebastian
Paust, Nils
Rombach, Markus
Lüddecke, Jan
Specht, Mara
Zengerle, Roland
Mitsakakis, Konstantinos
author_facet Hin, Sebastian
Paust, Nils
Rombach, Markus
Lüddecke, Jan
Specht, Mara
Zengerle, Roland
Mitsakakis, Konstantinos
author_sort Hin, Sebastian
collection PubMed
description Centrifugal microfluidics enables fully automated molecular diagnostics at the point-of-need. However, the integration of solid-phase nucleic acid extraction remains a challenge. Under this scope, we developed the magnetophoresis under continuous rotation for magnetic bead-based nucleic acid extraction. Four stationary permanent magnets are arranged above a cartridge, creating a magnetic field that enables the beads to be transported between the chambers of the extraction module under continuous rotation. The centrifugal force is maintained to avoid uncontrolled spreading of liquids. We concluded that below a frequency of 5 Hz, magnetic beads move radially inwards. In support of magnetophoresis, bead inertia and passive geometrical design features allow to control the azimuthal bead movement between chambers. We then demonstrated ferrimagnetic bead transfer in liquids with broad range of surface tension and density values. Furthermore, we extracted nucleic acids from lysed Anopheles gambiae mosquitoes reaching comparable results of eluate purity (LabDisk: A260/A280 = 1.6 ± 0.04; Reference: 1.8 ± 0.17), and RT-PCR of extracted RNA (LabDisk: Ct = 17.9 ± 1.6; Reference: Ct = 19.3 ± 1.7). Conclusively, magnetophoresis at continuous rotation enables easy cartridge integration and nucleic acid extraction at the point-of-need with high yield and purity.
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spelling pubmed-97875632022-12-24 Magnetophoresis in Centrifugal Microfluidics at Continuous Rotation for Nucleic Acid Extraction Hin, Sebastian Paust, Nils Rombach, Markus Lüddecke, Jan Specht, Mara Zengerle, Roland Mitsakakis, Konstantinos Micromachines (Basel) Article Centrifugal microfluidics enables fully automated molecular diagnostics at the point-of-need. However, the integration of solid-phase nucleic acid extraction remains a challenge. Under this scope, we developed the magnetophoresis under continuous rotation for magnetic bead-based nucleic acid extraction. Four stationary permanent magnets are arranged above a cartridge, creating a magnetic field that enables the beads to be transported between the chambers of the extraction module under continuous rotation. The centrifugal force is maintained to avoid uncontrolled spreading of liquids. We concluded that below a frequency of 5 Hz, magnetic beads move radially inwards. In support of magnetophoresis, bead inertia and passive geometrical design features allow to control the azimuthal bead movement between chambers. We then demonstrated ferrimagnetic bead transfer in liquids with broad range of surface tension and density values. Furthermore, we extracted nucleic acids from lysed Anopheles gambiae mosquitoes reaching comparable results of eluate purity (LabDisk: A260/A280 = 1.6 ± 0.04; Reference: 1.8 ± 0.17), and RT-PCR of extracted RNA (LabDisk: Ct = 17.9 ± 1.6; Reference: Ct = 19.3 ± 1.7). Conclusively, magnetophoresis at continuous rotation enables easy cartridge integration and nucleic acid extraction at the point-of-need with high yield and purity. MDPI 2022-11-29 /pmc/articles/PMC9787563/ /pubmed/36557411 http://dx.doi.org/10.3390/mi13122112 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Hin, Sebastian
Paust, Nils
Rombach, Markus
Lüddecke, Jan
Specht, Mara
Zengerle, Roland
Mitsakakis, Konstantinos
Magnetophoresis in Centrifugal Microfluidics at Continuous Rotation for Nucleic Acid Extraction
title Magnetophoresis in Centrifugal Microfluidics at Continuous Rotation for Nucleic Acid Extraction
title_full Magnetophoresis in Centrifugal Microfluidics at Continuous Rotation for Nucleic Acid Extraction
title_fullStr Magnetophoresis in Centrifugal Microfluidics at Continuous Rotation for Nucleic Acid Extraction
title_full_unstemmed Magnetophoresis in Centrifugal Microfluidics at Continuous Rotation for Nucleic Acid Extraction
title_short Magnetophoresis in Centrifugal Microfluidics at Continuous Rotation for Nucleic Acid Extraction
title_sort magnetophoresis in centrifugal microfluidics at continuous rotation for nucleic acid extraction
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9787563/
https://www.ncbi.nlm.nih.gov/pubmed/36557411
http://dx.doi.org/10.3390/mi13122112
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