title: Exiqon miRCURY LNA microRNA array v11.0 (miRBase 14.0)
status: Public on Feb 01 2013
contact.city: Denver
contact.first: Scott
contact.middle: A
contact.last: Barbee
contact.email: scott.barbee@du.edu
contact.state: CO
contact.address: 2190 E. ILIFF Ave.
contact.department: Biological Sciences
contact.country: USA
contact.institute: University of Denver
contact.phone: 303-871-3582
organism: Drosophila melanogaster
manufacture_protocol: The quality of the total RNA was verified by an Agilent 2100 Bioanalyzer profile. 0.5 µg total RNA from sample and reference was labelled with Hy3™ and Hy5™ fluorescent label, respectively, using the miRCURY™ LNA Array power labelling kit (Exiqon, Denmark) following the procedure described by the manufacturer. The Hy3™-labeled samples and a Hy5™-labeled reference RNA sample were mixed pair-wise and hybridized to the miRCURY™ LNA array version 11.0 Other Species (Exiqon, Denmark), which contains capture probes targeting all miRNAs for all other species than human, mouse or rat registered in the miRBASE version 14.0 at the Sanger Institute. The hybridization was performed according to the miRCURY™ LNA array manual using a Tecan HS4800 hybridization station (Tecan, Austria). After hybridization the microarray slides were scanned and stored in an ozone free environment (ozone level below 2.0 ppb) in order to prevent potential bleaching of the fluorescent dyes. The miRCURY™ LNA array microarray slides were scanned using the Agilent G2565BA Microarray Scanner System (Agilent Technologies, Inc., USA) and the image analysis was carried out using the ImaGene 8.0 software (BioDiscovery, Inc., USA). The quantified signals were background corrected (Normexp with offset value 10 – Ritchie et al., 2007) and normalized using the global Lowess (LOcally WEighted Scatterplot Smoothing) regression algorithm.