Chapter title |
Application of Individual qPCR Performance Parameters for Quality Control of Circulating MicroRNA Data
|
---|---|
Chapter number | 14 |
Book title |
MicroRNA and Cancer
|
Published in |
Methods in molecular biology, January 2018
|
DOI | 10.1007/978-1-4939-7435-1_14 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7433-7, 978-1-4939-7435-1
|
Authors |
Anna Brunet-Vega, María Elisa Quílez, María José Ramírez-Lázaro, Sergio Lario, Brunet-Vega, Anna, Quílez, María Elisa, Ramírez-Lázaro, María José, Lario, Sergio |
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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