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Highly Sensitive microRNA Array Performance

Fluorescent dyes or other detectable moieties can be appended to the amine-modified miRNAs using NHS-ester derivatized dyes. The mixture of modified and unmodified nucleotides improves tailing efficiency and reduces fluorescence quenching that occurs when dye molecules are located on adjacent or nearby nucleotides.

Together the mirVana miRNA Labeling Kit and mirVana miRNA Probe Set provide the best combination of sensitivity and accuracy of any procedure for miRNA profiling. Here we validate key assay characteristics including sensitivity, specificity, and accuracy of this miRNA microarray procedure.


Highly Sensitive Limit of Detection

To determine sensitivity of the mirVana miRNA array system, we isolated the miRNA fraction from liver RNA samples then spiked in various amounts of miR-124, a brain-specific miRNA. Liver miRNA samples with and without added miR-124 were then labeled using the 3' tailing method, and each miR-124-containing sample was cohybridized with a differentially labeled liver miRNA sample missing miR-124. Analysis of raw signal intensities and the Log2(R/G Normalized Ratio) (Mean) of miR-124 expression indicated that signal significantly above slide background was detectable with femtomole amounts of miR-124 (Figure 1). This level of detection represents as little as 0.1% of the overall miRNA population in a 10 g total RNA sample.

Figure 1. miRNA Array Sensitivity. Raw intensities of miR-124 expression measured from liver miRNA samples with
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