Solvent-selective routing for centrifugally automated solid-phase purification of RNA

被引:0
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作者
Nikolay Dimov
Eoin Clancy
Jennifer Gaughran
David Boyle
Darren Mc Auley
Macdara T. Glynn
Róisín M. Dwyer
Helena Coughlan
Thomas Barry
Louise M. Barrett
Terry J. Smith
Jens Ducrée
机构
[1] Dublin City University,National Centre of Sensor Research, School of Physical Sciences, Biomedical Diagnostics Institute
[2] University College London,Department of Advanced Biochemical Engineering
[3] National University of Ireland,National Centre for Biomedical Engineering Science, School of Natural Sciences, Biomedical Diagnostics Institute Programme
[4] National University of Ireland,School of Medicine
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关键词
Lab-on-a-disc; RNA purification; Solvent-selective valves; Molecular diagnostics;
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摘要
We present a disc-based module for rotationally controlled solid-phase purification of RNA from cell lysate. To this end, multi-stage routing of a sequence of aqueous and organic liquids into designated waste and elution reservoirs is implemented by a network of strategically placed, solvent-selective composite valves . Using a bead-based stationary phase at the entrance of the router, we show that total RNA is purified with high integrity from cultured MCF7 and T47D cell lines, human leucocytes and Haemophilus influenzae cell lysates. Furthermore, we demonstrate the broad applicability of the device through the in vitro amplification of RNA purified on-disc using RT-PCR and NASBA. Our novel router will be at the pivot of a forthcoming, fully integrated and automated sample preparation system for RNA-based analysis .
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页码:859 / 871
页数:12
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