Quantification of miRNA precursors in Mammalian Cells

Martin Kreutz QIAGEN GmbH, Germany

Abstract
With the miScript system Qiagen has developed a robust and accurate method for transcriptome-wide miRNA quantification using SYBR Green detection-based, real-time PCR. The system allows the detection of mature miRNAs as well other short, non-coding RNAs and even mRNAs from the same cDNA. Mature miRNAs are processed from longer precursor molecules, the so called pri-miRNAs and pre-miRNAs. These precursors are first transcribed in the nucleus and then further processed in nucleus and cytosol. Recent reports of the regulation of mature miRNA expression through precursor processing have led to an increased interest in the expression patterns of these miRNA precursors. Here we demonstrate how both mature miRNAs and miRNA precursors can be specifically detected from the same cDNA sample. A new tool for functional analysis of the interaction of miRNAs and their targets will also be discussed.