Can a TaqMan Pri-miRNA Assay detect more than a single stem-loop?
TaqMan Pri-miRNA Assays are designed to detect the transcript that contains the targeted stem loop. In some instances, e.g. when stem-loop sequences are clustered together and are transcribed on the same transcript, a TaqMan Pri-miRNA Assay targeting one of the stem-loops will also detect the other stem loops in the cluster.
Do the TaqMan MicroRNA Assays detect only the mature form of a miRNA? Vice versa, do the TaqMan Pri-miRNA Assays detect only the pri-form?
Yes. The specific stem-loop chemistry of the TaqMan MicroRNA Assay and its design prevent amplification of the pri-form. Vice versa, the chemistry of the TaqMan Pri-miRNA Assays is not amenable to amplify the mature form. In particular cases, assays designed to detect mature miRNAs derived from the 3' arm of the stem loop may also detect pre-miRNA.
What endogenous control to use for pri-miRNA?
TaqMan endogenous control assays are compatible for pri-miRNA (visit the TaqMan Endogenous Control Assays web portal for more information).
Which kit should I use for the reverse transcription step for TaqMan Pri-miRNA assays (Cat. No. 4427012)?