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AAT Bioquest

What methods can be used to study miRNA function?

Posted March 18, 2024


Answer

Traditional methods to study miRNA function include northern blot, microarray analysis, in situ-hybridization, and quantitative PCR. 

  • Quantitative PCR (qPCR) is used for detecting miRNAs present at low levels in biological samples. It is highly sensitive and widely used for its ability to precisely quantify gene expression differences between samples. In qPCR, the abundance of a specific miRNA is measured by quantifying the amplification of its complementary DNA (cDNA) generated through reverse transcription. 
  • Northern blotting is a traditional method for detecting and quantifying RNA molecules, including microRNAs. In this technique, RNA samples are separated by gel electrophoresis, transferred to a membrane, and hybridized with microRNA-specific probes. The resulting bands are visualized using chemiluminescence, providing information on microRNA size, abundance, and expression patterns. 
  • Microarray analysis allows for high-throughput profiling of microRNA expression levels across thousands of microRNAs simultaneously. In this method, microRNA molecules are labeled with fluorescent dyes or biotin and hybridized to microarray chips containing complementary probes. The intensity of fluorescence or signal generated upon hybridization reflects the abundance of each microRNA in the sample. 
  • In situ hybridization is a technique for visualizing the cellular localization of microRNAs within tissues or cells. It involves hybridizing fluorescently labeled microRNA probes to fixed tissue sections or cultured cells, followed by microscopy-based detection of probe binding.
Additional resources

Gene Expression Is Analyzed by Tracking RNA

DNA and RNA Quantitation

Portelite™ Fluorimetric RNA Quantitation Kit*Optimized for Cytocite™ and Qubit™ Fluorometers*