microRNA Quantification by qRT-PCR

microRNA qRT-PCR kits with specific RT mixes and validated miRNA primer sets

Specialised qRT-PCR kits for microRNA quantification differ from mRNA qPCR because mature miRNAs are short (22 nt) and require either stem-loop primed reverse transcription or polyA-tailing-plus-universal-primer strategies. The category includes miRNA-specific RT mixes, qPCR master mixes, and primer or probe sets against thousands of validated miRNA targets.

Pick by chemistry and normalisation.

1. RT chemistry. Stem-loop primed gives highest specificity for the specific miRNA isoform. PolyA-tailed universal RT gives broader coverage with one cDNA pool.
2. Panel coverage. Validated miRNAs above 2,000 across human, mouse, rat is standard.
3. Normalisation. Use multiple stable reference miRNAs or invariant small nucleolar RNAs. Never normalise to a single miRNA in circulating biomarker studies.
4. Input. Some kits accept exosomal RNA at picogram input.
5. Caveat. Normalisation strategy is the single biggest source of contradictory miRNA results in the literature. Pick early and lock the protocol.

SBI System Biosciences for exosome-focused miRNA quantification. GeneCopoeia for stem-loop primer panels for cell-based miRNA analysis.