Pri-miRNA Analysis Using Real-Time PCR
How It Works
MicroRNA “genes” are transcribed in the nucleus creating a long pri-miRNA transcript. The pri-miRNA sequence contains one or more stem-loop sequences, each containing mature miRNA sequences. TaqMan® Pri-miRNA Assays are designed in close proximity to each independent stem-loop sequence identified in the Sanger miRBase sequence repository (http://www.mirbase.org) using Applied Biosystems’ state-of-the-art bioinformatics pipeline. This approach ensures that the primary transcript containing the stem-loop of interest is being measured.
Pri-miRNA Analysis Experimental Workflow
Life Technologies offers a variety of kits that preserve small RNAs such as miRNAs. While it is not essential to use them due to the larger size of primary miRNA transcripts, these kits do enable both pri-miRNA and mature miRNA quantitation from the same RNA sample. Each kit is ideal for miRNA analysis because they enable quantitative recovery of all RNA >10nt in length and maintenance of representative amounts of all RNA (eliminating experimental bias). To ensure accurate quantitation with TaqMan® Pri-miRNA Assays, samples must be free from genomic DNA.
TaqMan® miRNA ABC Purification Kit
mirVana™ miRNA Isolation Kit
RecoverAll™ Total Nucleic Acid Isolation Kit for FFPE
TURBO DNA-free™ Kit
TaqMan® MicroRNA Cells-to-CT™ Kit