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Detection of microRNAs Using Chip-Based QuantStudio 3D Digital PCR

  • Cristina Borzi
  • Linda Calzolari
  • Davide Conte
  • Gabriella SozziEmail author
  • Orazio Fortunato
Part of the Methods in Molecular Biology book series (MIMB, volume 1580)

Abstract

Digital PCR (dPCR) is an innovative approach for detection and quantification of nucleic acid that offers an alternative method to conventional real-time quantitative PCR for absolute quantification. dPCR is a highly precise and sensitive technique that does not require a standard reference, making it a suitable method for the detection of microRNAs. The potential of these small noncoding RNA as biomarkers is on the rise, especially due to their presence in body fluids, making them easily accessible. Nevertheless, the problem of lack of consensus regarding an optimal method for miRNAs normalization compromises their use. Here, we describe an innovative method for the absolute quantification of miRNAs across different types of biological samples using a chip-based platform, the QuantStudio 3D digital PCR.

Key words

Digital PCR Absolute quantification miRNAs 

Notes

Acknowledgments

This work was supported by AIRC (Associazione Italiana per la Ricerca sul Cancro): IG14318 to G.S., and 12162 (Special Program “Innovative Tools for Cancer Risk Assessment and early Diagnosis,” 5 × 1000); Italian Ministry of Health RF-2010-2306232 to G.S.

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Cristina Borzi
    • 1
  • Linda Calzolari
    • 1
  • Davide Conte
    • 1
  • Gabriella Sozzi
    • 1
    Email author
  • Orazio Fortunato
    • 1
  1. 1.Tumor Genomics Unit, Department of Experimental Oncology and Molecular MedicineFondazione IRCCS Istituto Nazionale dei TumoriMilanItaly

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