Total RNA was prepared using Trizol Reagent according to the manufacturer’s instructions (Invitrogen) and was further purified using RNeasy columns (Qiagen).
Label
Biotin
Label protocol
Seven μg of total RNA were subjected to One-Cycle Target Labeling Assay (Affymetrix) to generate biotinylated cRNA targets for hybridization to Affymetrix U133A 2.0 microarray.
Hybridization protocol
Standard hybridization to Affymetrix U133A 2.0 microarray.
Scan protocol
Standard scanning protocol using GeneChip Scanner.
Description
N/A
Data processing
The raw data were processed by Affymetrix GCOS software to obtain detection calls and signal values, and then normalized by scaling. Signals were then used in further analysis, namely batch effect adjusted and log2 median-normalized using COMBAT R.
