Mircury lna array power labeling kit

The experiments using the Exiqon platform were performed by Exiqon (Vedbaek, Denmark). Briefly, total RNA samples (0.75 μg) and reference RNA samples were labeled with Hy3™ and Hy5™ fluorescent labels, respectively, using the miRCURY LNA™ Array power labeling kit (Exiqon), following the procedures described by the manufacturer. No dye swap experiments were performed. Each Hy3™-labeled sample was mixed with the Hy5™-labeled reference RNA sample and then hybridized on the miRCURY LNA™ miRNA Arrays (v.11.0) following the “common reference design” [35 (link)]. For each tissue type (lung and blood), the reference RNA consisted in a mixture of RNA extracted from all the samples in the experiment. The miRCURY array contains a number of capture probes for hybridization of a range of synthetic spike-in controls. The spike-in controls “a”, “b”, “c”, “d”, “e”, “f”, “g”, “h”, “i”, and “j” were spiked into the labeling reaction at various concentrations. Hybridization was performed according to the miRCURY LNA™ array manual. Following hybridization and processing, the arrays were scanned using the Agilent G2565BA Microarray Scanner System (Agilent Technologies, Inc., Santa Clara, CA) and image analysis was performed using the ImaGene 8.0 software (BioDiscovery, Inc., Santa Clara, CA) to quantify the signals. The raw data were provided by Exiqon as ImaGene text files.