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Microfluidics and Nanofluidics

, Volume 18, Issue 5–6, pp 859–871 | Cite as

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 Paper

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 .

Keywords

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

Notes

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.

Supplementary material

Supplementary material 1 (mpg 23830 KB)

10404_2014_1477_MOESM2_ESM.pdf (868 kb)
Supplementary material 2 (pdf 867 KB)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  • Nikolay Dimov
    • 1
    • 2
  • Eoin Clancy
    • 3
  • Jennifer Gaughran
    • 1
  • David Boyle
    • 1
  • Darren Mc Auley
    • 1
  • Macdara T. Glynn
    • 1
  • Róisín M. Dwyer
    • 4
  • Helena Coughlan
    • 3
  • Thomas Barry
    • 3
  • Louise M. Barrett
    • 1
  • Terry J. Smith
    • 3
  • Jens Ducrée
    • 1
  1. 1.National Centre of Sensor Research, School of Physical Sciences, Biomedical Diagnostics InstituteDublin City UniversityGlasnevin, Dublin 9Ireland
  2. 2.Department of Advanced Biochemical EngineeringUniversity College LondonLondonUK
  3. 3.National Centre for Biomedical Engineering Science, School of Natural Sciences, Biomedical Diagnostics Institute ProgrammeNational University of IrelandGalwayIreland
  4. 4.School of MedicineNational University of IrelandGalwayIreland

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