In the paper Two-tailed RT-qPCR panel for quality control of circulating microRNA studies published in Scientific Reports from Nature, experts in the field present an RT-qPCR based protocol to assess the technical performance of workflows for analysis of microRNAs in body fluids such as serum and plasma.
Despite several advances, circulating microRNA research has been hampered by inconsistency and poor reproducibility. The Two-tailed RT-qPCR quality control panel developed in the report is a simple yet powerful tool for researchers to generally improve the reliability of circulating microRNA data.
Abstract (from Nature):
Circulating cell-free microRNAs are promising candidates for minimally invasive clinical biomarkers for the diagnosis, prognosis and monitoring of many human diseases. Despite substantial efforts invested in the field, the research so far has failed to deliver expected results.
One of the contributing factors is general lack of agreement between various studies, partly due to the considerable technical challenges accompanying the workflow. Pre-analytical variables including sample collection, RNA isolation, and quantification are sources of bias that may hamper biological interpretation of the results.
Here, we present a Two-tailed RT-qPCR panel for quality control, monitoring of technical performance, and optimization of microRNA profiling experiments from biofluid samples. The Two-tailed QC (quality control) panel is based on two sets of synthetic spike-in molecules and three endogenous microRNAs that are quantified with the highly specific Two-tailed RT-qPCR technology, developed by TATAA Biocenter.
The QC panel is a cost-effective way to assess quality of isolated microRNA, degree of inhibition, and erythrocyte contamination to ensure technical soundness of the obtained results. We provide assay sequences, detailed experimental protocol and guide to data interpretation. The application of the QC panel is demonstrated on the optimization of RNA isolation from biofluids with the miRNeasy Serum/Plasma Advanced Kit (Qiagen).
