Have these miRIDIAN micrRNA Mimics and Hairpin Inhibitors products been tested successfully by your company? Can I use them directly without testing?
We have tested miRIDIAN micrRNA Mimics and Hairpin Inhibitors for specific microRNA activity using a reporter plasmid-based system in a few cell lines. We have tested them in the low nanomolar range (50 nM or less) in a luciferase reporter system; however, ideal concentrations MUST be empirically determined for each microRNA inhibitor and mimic tested in each cell type. We always conduct dose and time curves. For each miRNA in each cell line, we suggest time points at 24, 48 and 72 hours and concentrations tested at 100 nM, 50 nM, 10 nM, 2 nM, 0.4 nM. Working concentrations of anywhere from 5-10 picomolar to 20 micromolar have been demonstrated. MicroRNAs can be highly expressed, lowly expressed, not expressed at all or work in cooperation with other microRNAs so you should titrate different amounts of mimics/inhibitors depending on the endogenous expression levels of the miRNAs and find, for each experiment, a proper working concentration. For additional information please download the Technical Note on using Mimics and Inhibitors for microRNA (miRNA) Functional Analysis.