TaqMan assays for quantifying precursor microRNA and mature microRNA. (a) TaqMan assay designed to quantify hairpin-containing microRNA precursors. RT reaction and PCR amplification are performed using gene-specific primers. Quantification is carried out using TaqMan minor groove binder (MGB) probes that are specific to the loop portion of the microRNA precursor. (b) TaqMan assay designed to quantify mature microRNA. This assay utilizes a target specific stem-loop, reverse transcription primer for RT reaction. The stem-loop structure provides specificity for only the mature microRNA target and forms a RT primer/mature microRNA-chimera that extends the end of the microRNA. The resulting RT product will be quantified using standard TaqMan quantitative real-time PCR method.
