Total RNA was isolated using the High Pure RNA Paraffin Kit (Roche, IN, USA) per manufacturer instructions.
Label
biotin
Label protocol
RNA was amplified and labeled using the SensationPlus™ Kit (Affymetrix, Santa Clara, CA, USA) per manufacturer instructions.
Hybridization protocol
Following fragmentation, 10 ug of cRNA were hybridized for 16 hr at 45C on HG-U133A 2.0 arrays. GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
Scan protocol
GeneChips were scanned using the Affymetrix GC3000 scanner.
Data processing
After scanning, the raw intensities for every probe were stored in electronic files (in .DAT and .CEL formats) by the Affymetrix GeneChip Operating Software (GCOSv1.4). Background correction, normalization, and estimation of probe set expression summaries was performed using the log-scale Robust Multiarray Analysis (RMA) method.
