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Continuous Microfluidic Purification of DNA Using Magnetophoresis

Automatic microfluidic purification of nucleic acid is predictable to reduce the input of original samples and improve the throughput of library preparation for sequencing. Here, we propose a novel microfluidic system using an external NdFeB magnet to isolate DNA from the polymerase chain reaction (...

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Detalles Bibliográficos
Autores principales: Xu, Ying, Zhang, Zhen, Su, Zhen, Zhou, Xiaoxiang, Han, Xiaoming, Liu, Quanjun
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7074667/
https://www.ncbi.nlm.nih.gov/pubmed/32054004
http://dx.doi.org/10.3390/mi11020187
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author Xu, Ying
Zhang, Zhen
Su, Zhen
Zhou, Xiaoxiang
Han, Xiaoming
Liu, Quanjun
author_facet Xu, Ying
Zhang, Zhen
Su, Zhen
Zhou, Xiaoxiang
Han, Xiaoming
Liu, Quanjun
author_sort Xu, Ying
collection PubMed
description Automatic microfluidic purification of nucleic acid is predictable to reduce the input of original samples and improve the throughput of library preparation for sequencing. Here, we propose a novel microfluidic system using an external NdFeB magnet to isolate DNA from the polymerase chain reaction (PCR) mixture. The DNA was purified and isolated when the DNA-carrying beads transported to the interface of multi-laminar flow under the influence of magnetic field. Prior to the DNA recovery experiments, COMSOL simulations were carried out to study the relationship between trajectory of beads and magnet positions as well as fluid velocities. Afterwards, the experiments to study the influence of varying velocities and input of samples on the DNA recovery were conducted. Compared to experimental results, the relative error of the final position of beads is less than 10%. The recovery efficiency decreases with increase of input or fluid velocity, and the maximum DNA recovery efficiency is 98.4% with input of l00 ng DNA at fluid velocity of 1.373 mm/s. The results show that simulations significantly reduce the time for parameter adjustment in experiments. In addition, this platform uses a basic two-layer chip to realize automatic DNA isolation without any other liquid switch value or magnet controller.
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spelling pubmed-70746672020-03-20 Continuous Microfluidic Purification of DNA Using Magnetophoresis Xu, Ying Zhang, Zhen Su, Zhen Zhou, Xiaoxiang Han, Xiaoming Liu, Quanjun Micromachines (Basel) Article Automatic microfluidic purification of nucleic acid is predictable to reduce the input of original samples and improve the throughput of library preparation for sequencing. Here, we propose a novel microfluidic system using an external NdFeB magnet to isolate DNA from the polymerase chain reaction (PCR) mixture. The DNA was purified and isolated when the DNA-carrying beads transported to the interface of multi-laminar flow under the influence of magnetic field. Prior to the DNA recovery experiments, COMSOL simulations were carried out to study the relationship between trajectory of beads and magnet positions as well as fluid velocities. Afterwards, the experiments to study the influence of varying velocities and input of samples on the DNA recovery were conducted. Compared to experimental results, the relative error of the final position of beads is less than 10%. The recovery efficiency decreases with increase of input or fluid velocity, and the maximum DNA recovery efficiency is 98.4% with input of l00 ng DNA at fluid velocity of 1.373 mm/s. The results show that simulations significantly reduce the time for parameter adjustment in experiments. In addition, this platform uses a basic two-layer chip to realize automatic DNA isolation without any other liquid switch value or magnet controller. MDPI 2020-02-11 /pmc/articles/PMC7074667/ /pubmed/32054004 http://dx.doi.org/10.3390/mi11020187 Text en © 2020 by the authors. 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 (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Xu, Ying
Zhang, Zhen
Su, Zhen
Zhou, Xiaoxiang
Han, Xiaoming
Liu, Quanjun
Continuous Microfluidic Purification of DNA Using Magnetophoresis
title Continuous Microfluidic Purification of DNA Using Magnetophoresis
title_full Continuous Microfluidic Purification of DNA Using Magnetophoresis
title_fullStr Continuous Microfluidic Purification of DNA Using Magnetophoresis
title_full_unstemmed Continuous Microfluidic Purification of DNA Using Magnetophoresis
title_short Continuous Microfluidic Purification of DNA Using Magnetophoresis
title_sort continuous microfluidic purification of dna using magnetophoresis
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7074667/
https://www.ncbi.nlm.nih.gov/pubmed/32054004
http://dx.doi.org/10.3390/mi11020187
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