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The same three sets of samples and one additional set of samples were analyzed using the Nanostring nCounter solution hybridization assay. Array hybridization and analyses MiRNA microarray analysis was performed by Genome Explorations. RNA was processed and labeled according to standard RT-IVT methods. Labeled cRNA (15.0 ?g) was fragmented by ion-mediated hydrolysis and hybridized for 16 h at 45��C to Affymetrix miRNA v2.0 array. The arrays were read and the log2 mean intensities were used for further analysis. Data were first filtered for human miRNA and miRNA probe sets with at least one detection call among the 18 samples were included for Perform Principal Components Analysis (PCA) to identify outlier samples. One 39.5��C sample was identified as an extreme outlier and removed from CP-868596 manufacturer further analysis. ANOVA was performed PTPRJ for all six groups (with and without Benjamini-Hochberg FDR correction). Independent t-tests were performed for each pairwise comparison. Data were filtered for ANOVA P values 0.585 (1.5-fold) and t-test P value learn more were hybridized to color-coded reporter probes and biotinylated capture probes, immobilized onto a cartridge, and each immobilized miRNA was identified based on its color code and quantified. A total of 811 human and human-associated viral miRNAs were simultaneously assayed. All assays were performed using 100 ng of total RNA following the standard nCounter miRNA Assay Protocol. Hybridizations were performed automatically by mixing 5 ?L of each miRNA multiplex assay with 20 ?L NanoString nCounter reporter probe mix and 5 ?L capture probe mix (30 ?L total reaction volume), and hybridizing at 65��C for 18 h. The raw miRNA counts were analyzed using the nSolver software and normalized using the top 100 normalization method. The normalized values were then subject to two-way ANOVA and individual pair wise t-tests. miRNA with ANOVA P values 1.5-fold and t-test P value