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Droplet Microfluidic Device Fabrication and Use for Isothermal Amplification and Detection of MicroRNA

  • Maria Chiara Giuffrida
  • Roberta D’Agata
  • Giuseppe SpotoEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1580)

Abstract

Droplet microfluidics combined with the isothermal circular strand displacement polymerization (ICSDP) represents a powerful new technique to detect both single-stranded DNA and microRNA sequences. The method here described helps in overcoming some drawbacks of the lately introduced droplet polymerase chain reaction (PCR) amplification when implemented in microfluidic devices. The method also allows the detection of nanoliter droplets of nucleic acids sequences solutions, with a particular attention to microRNA sequences that are detected at the picomolar level. The integration of the ICSDP amplification protocol in droplet microfluidic devices reduces the time of analysis and the amount of sample required. In addition, there is also the possibility to design parallel analyses to be integrated in portable devices.

Key words

Circular strand displacement polymerization Isothermal amplification Droplet microfluidics MicroRNA Nucleic acids amplification 

Notes

Acknowledgments

MIUR (PRIN 20093N774P) and Ministry of Health, Italy (n. 098/GR-2009-1596647), are acknowledged for partial financial support.

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Maria Chiara Giuffrida
    • 1
  • Roberta D’Agata
    • 1
  • Giuseppe Spoto
    • 1
    • 2
    Email author
  1. 1.I.N.B.B. ConsortiumRomeItaly
  2. 2.Dipartimento di Scienze ChimicheUniversità di CataniaCataniaItaly

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