tissue: eosinophilic CRSsNP from sinonasal mucosa disease state: CRSsNP
Extracted molecule
total RNA
Extraction protocol
Total RNA was extracted using TRIzol® Reagent (Invitrogen) according to the manufacturer’s protocol.
Label
Hy5
Label protocol
2 μg of total RNA from each sample was fluorochrome labeled Exiqon®, Vedbaek, Denmark using the miRCURY™ LNA microRNA Power labeling kit.
Hybridization protocol
The Hy5™ fluorescent labeled miRNAs were hybridized to Exiqon miRCURY locked nucleic acid (LNA) array (version 9.2) (Exiqon, Vedbaek, Denmark) at 52ºC for overlight.
Scan protocol
Arrays were scanned, after hybridization and washing, using the G2505B scanning system (Agilent®, Santa Clara, CA, USA) by Exiqon® (Vedbaek, Denmark).
Data processing
The intensity of green signal is calculated after background subtraction and replicated spots on the same slide have been averaged by getting a median intensity. We use Median Normalization Method to obtain “normalized data”, the median is 50 percent quantile of microRNA intensity which is larger than 50 in all samples after background correction.
