Related Experiment Video
Updated: Jul 16, 2026

Probe-based Real-time PCR Approaches for Quantitative Measurement of microRNAs
Published on: April 14, 2015
RNA interference: PCR strategies for the quantification of stable degradation-fragments derived from siRNA-targeted
Peter Hahn1, Cornelia Schmidt, Martin Weber
1QIAGEN GmbH, QIAGEN Strasse 1, D-40724 Hilden, Germany.
Abstract:
mRNA targeted by siRNA is endogeneously cleaved into a 5'- and a 3'-fragment and finally degraded in cells. Little is known about the relative stability and degradation kinetics of these 5'- and 3'-fragments after the siRNA mediated first cut. We present a qRT-PCR protocol which allows the determination of the optimal time point for mRNA analyses, helping to avoid the generation of false positive effects in downstream experiments, such as microarray analysis, which may be caused by undegraded fragments of a siRNA-targeted mRNA.
Related Concept Videos
RNA Interference
This process occurs naturally in cells, often through the activity of genomically-encoded microRNAs. Researchers can take advantage of this mechanism by introducing synthetic RNAs to deactivate specific genes for research or therapeutic purposes. For example, RNAi could be used...
Nonsense-mediated mRNA Decay
Usually, Upf3 binds to an Exon Junction Complex (EJC) at mRNA splice sites. If a ribosome fully translates the mRNA,...
Real Time RT-PCR
The real-time quantification of the number of amplified products is...
Experimental RNAi

