Trizol extraction of total RNA was performed according to the manufacturer's instructions.
Label
Biotin
Label protocol
Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 15 microg total RNA.
Hybridization protocol
15 ug of total RNA was reverse-transcribed with oligo-dT primer containing a T7 RNA polymerase-binding site. An in vitro transcription was performed on this cDNA. DNA from the samples was fragmented in 40 mM Tris acetate, 100 mM potassium acetate, and 30 mM MgCl2 (pH 8.1) at 95°C to reduce secondary structures. 15 ug of cRNA were hybridized to an Affymetrix HuGeneFL microarray, washed and stained.