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DNA purification using dynamic solid-phase extraction on a rotationally-driven polyethylene-terephthalate microdevice

Authors :
R. Aranda
M. Meija
Juliane C. Borba
James P. Landers
Doris M. Haverstick
Gavin T. Garner
Daniel L. Mills
Kimberly Jackson
Emanuel Carrilho
K.E. Olson
Source :
Repositório Institucional da USP (Biblioteca Digital da Produção Intelectual), Universidade de São Paulo (USP), instacron:USP
Publication Year :
2016
Publisher :
Elsevier BV, 2016.

Abstract

We report the development of a disposable polyester toner centrifugal device for semi-automated, dynamic solid phase DNA extraction (dSPE) from whole blood samples. The integration of a novel adhesive and hydrophobic valving with a simple and low cost microfabrication method allowed for sequential addition of reagents without the need for external equipment for fluid flow control. The spin-dSPE method yielded an average extraction efficiency of ∼45% from 0.6 μL of whole blood. The device performed single sample extractions or accommodate up to four samples for simultaneous DNA extraction, with PCR-readiness DNA confirmed by effective amplification of a β-globin gene. The purity of the DNA was challenged by a multiplex amplification with 16 targeted amplification sites. Successful multiplexed amplification could routinely be obtained using the purified DNA collected post an on-chip extraction, with the results comparable to those obtained with commercial DNA extraction methods. This proof-of-principle work represents a significant step towards a fully-automated low cost DNA extraction device.

Details

ISSN :
00032670
Volume :
937
Database :
OpenAIRE
Journal :
Analytica Chimica Acta
Accession number :
edsair.doi.dedup.....dd8aa496cdeb3a61b214d8b78761e80e
Full Text :
https://doi.org/10.1016/j.aca.2016.06.036