An Enhanced Quantitative Reverse Transcription-PCR Approach for Measuring Circulating MicroRNAs

Short (22 nt) RNAs called microRNAs (miRNAs) bind to and inhibit target messenger RNAs in gene regulatory networks. Recent study suggests that miRNAs circulate in a stable, cell-free form and that particular miRNAs in plasma or serum may be biomarkers for cancer and other diseases. Circulating miRNAs as biomarkers provide distinct challenges including pre-analytic variance and data standardization. We describe our qRT-PCR approach for measuring circulating miRNAs as biomarkers, as well as sample preparation, experimental design, and data processing issues.