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.
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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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Brunet-Vega, A., Quílez, M.E., Ramírez-Lázaro, M.J., Lario, S. (2018). Application of Individual qPCR Performance Parameters for Quality Control of Circulating MicroRNA Data. In: Wu, W. (eds) MicroRNA and Cancer. Methods in Molecular Biology, vol 1699. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7435-1_14
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DOI: https://doi.org/10.1007/978-1-4939-7435-1_14
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