Your TaqMan® MicroRNA Assays do not work

Common reasons for failure of miRNA detection are ranked below:

  • Inappropriate RNA Isolation methodology. Make sure that your RNA isolation method retains the miRNA fraction. Use the interactive RNA Isolation Decision Tree to select the right kit for your application.
  • Updates to miRNA sequence information.  Note that the registry for miRNA sequences, miRBase (http://www.mirbase.org), is constantly updated. These updates may include name, accession number, and even sequence changes of miRNAs. Therefore, the most effective way of locating the correct TaqMan® MicroRNA Assay on the Applied Biosystems website is to input the miRNA sequence into the search box under the Assay Search tab. Each assay design is connected to a specific miRNA sequence and will not be changed.

Tips/Tricks:

Use the mirVana™ miRNA Reference Panel (based on miRBase version 9.1), which contains a mixture of synthetic RNA oligonucleotides, as a positive control in real-time PCR. The complete annotation file of the reference panel is available here.