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

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

Research output: Contribution to journalArticlepeer-review

8 Citations (Scopus)
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Abstract

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.

Original languageEnglish
Pages (from-to)859-871
Number of pages13
JournalMicrofluidics and Nanofluidics
Volume18
Issue number5-6
Early online date12 Sept 2014
DOIs
Publication statusPublished - 1 May 2015

Keywords

  • Lab-on-a-disc
  • Molecular diagnostics
  • RNA purification
  • Solvent-selective valves

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