MiRNA Detection

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.