Microarray Analysis of miRNA in AD

1000 ng of RNA was labeled with Affymetrix FlashTag Biotin HSR RNA Labeling Kit (Affymetrix, Santa Clara, CA, USA) per the manufacturer’s instructions. Labeled RNA was hybridized to GeneChip microRNA 4.0 arrays (Affymetrix, Santa Clara, CA, USA). Chips were run using a Fluidics Station 450 Protocol (Affymetrix, Santa Clara, CA, USA). The results were exported to Partek Genomics Suite (version 6.6, Partek, St. Louis, MO, USA) for analysis. For initial comparison, the entire dataset was normalized using RMA-DABG in Expression Console, and data were exported to Partek Genomics Suite. Using this data, the mean probe intensities for each of the 2578 mature miRNA probes in the CSF and serum were plotted for all paired samples (n = 21). Next, the raw data from Expression console was exported, and normalization was carried out within each biofluid using RMA-DABG. Mature human miRNAs were analyzed and compared between disease groups in Partek Genomics Suite. ANOVA was used to compare miRNAs between patients with AD and controls. Serum and CSF were analyzed separately [49 (link)].