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Application of Individual qPCR Performance Parameters for Quality Control of Circulating MicroRNA Data

  • Anna Brunet-Vega
  • María Elisa Quílez
  • María José Ramírez-Lázaro
  • Sergio LarioEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1699)

Abstract

Although circulating miRNAs are promising candidates for biomarkers, several challenges must be overcome before miRNAs can be used for diagnosis and monitoring. One is quality control for the RNA extraction and quantification process. RNA quality control techniques are unsuitable as circulating miRNAs are in the fM range. Additionally, biofluids may contain inhibitors of the reverse transcriptase and polymerase enzymes, which may survive RNA purification. Herein, we describe the protocol we have used to check the robustness of miRNA purification and measurement by the addition of spike-ins and by evaluating the quality of the qPCR data, respectively.

Key words

Circulating miRNA Quantitative PCR Quality control PCR efficiency 

Notes

Acknowledgments

This study was supported by grant 1007/C/2013 from the Marató de TV3 (http://www.tv3.cat/marato/en/#), by grant PI12/01802 from Proyectos de Investigación en Salud, Instituto de Salud Carlos III (http://www.eng.isciii.es/ISCIII/es/general/index.shtml), and by grant CIR2011040 from the Fundació Parc Taulí (CIR2011040, http://www.tauli.cat/tauli/en/Fpt/fpt.htm). CIBERehd is funded by the Instituto de Salud Carlos III.

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Copyright information

© Springer Science+Business Media LLC 2018

Authors and Affiliations

  • Anna Brunet-Vega
    • 1
    • 3
  • María Elisa Quílez
    • 1
    • 2
    • 3
  • María José Ramírez-Lázaro
    • 2
    • 4
  • Sergio Lario
    • 2
    • 3
    • 4
    Email author
  1. 1.Oncology Service, Hospital de Sabadell, Corporació Sanitària Parc TaulíInstitut Universitari Parc Taulí-UABSabadellSpain
  2. 2.Digestive Diseases Service, Hospital de Sabadell, Corporació Sanitària Parc TaulíInstitut Universitari Parc Taulí-UABSabadellSpain
  3. 3.Fundació Parc Taulí, Corporació Sanitària Parc TaulíInstitut Universitari Parc Taulí-UABSabadellSpain
  4. 4.Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd)Instituto de Salud Carlos IIIMadridSpain

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