Total RNA from each time point was extracted using Trizol (Invitrogen) and purified using RNeasy columns (Qiagen, Valencia, CA, USA) according to the manufacturers’ protocol. After processing with DNase digestion and clean-up procedures, RNA samples were quantified, aliquoted, and stored at −80 °C until use. For quality control, RNA purity and integrity were evaluated by denaturing gel electrophoresis (OD 260/280 ratio) and analyzed on an Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA, USA).
Label
biotin
Label protocol
Biotinylated cRNA were prepared from 0.55 ug total RNA using the Illumina TotalPrep RNA Amplification Kit (Ambion, Austin,TX)
Hybridization protocol
Following fragmentation, 0.75 ug of cRNA were hybridized to the Illumina Expression Beadchip according to the protocols provided by the manufacturer
Scan protocol
Arrays were scanned using the Illumina Bead Array Reader Confocal Scanner
Data processing
Array data export processing and analysis was performed using Illumina GenomeStudio v2011.1 (Gene Expression Module v1.9.0)
