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

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 .

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Acknowledgments

This work was supported by the Science Foundation Ireland under Grant 10/CE/B1821. Jennifer Gaughran was funded through BioAnalysis and Therapeutics Structured Ph.D. Programme (Bio-AT) by HEA-PRTLI V. The authors also thank Prof. Richard O’Kennedy’s group in Dublin City University for allowing us to use their cell culture facilities. We would also like to thank Karsten Holona and her team in Harke PackPro for providing dissolvable films.

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Correspondence to Nikolay Dimov.

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Dimov, N., Clancy, E., Gaughran, J. et al. Solvent-selective routing for centrifugally automated solid-phase purification of RNA. Microfluid Nanofluid 18, 859–871 (2015). https://doi.org/10.1007/s10404-014-1477-9

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Keywords

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