Two methods were used to extract total RNA. One is by standard Trizol method and Second is by AllPrep DNA/RNA/Protein Mini Kit from QIAGEN. After extractiong, all are followed with qualification check by Agilent 2100.
Label
Biotin
Label protocol
As per manufacturer (Affymetrix).
Hybridization protocol
For mRNA array, total RNA was reverse transcripted to cDNA, reverse transcripted to labelled aRNA, purified, fragmented, and hybridized to each array chip according to the manufacturer's instructions.
Scan protocol
The mRNA arrays were washed using Affymetrix fluidics stations, and scanned using the Affymetrix GeneChip Scanner 3000.
Description
frozen tissue extracted by Trizol
Data processing
Microarray data were normalized using the RMA method .
