Total RNA was extracted from frozen tissue with a polytron homogenizer (Brinkman, Westbury, NY) and Trizol reagent (Invitrogen, Rockville, MD), and purified with an RNAse kit (Qiagen, Santa Clara, CA).
Label
Biotin
Label protocol
Total RNA was used as a template for double-stranded cDNA synthesis (Superscript Double-Stranded cDNA synthesis kit, Invitrogen, Carlsbad, CA). Biotin-labeledcDNA was synthesized (EnzoBioarray High Yield RNA transcription Labeling Kit, Affymetrix).
Hybridization protocol
Biotin-labeled cDNA was hybridized to Affymetrix Human Genome U133 Plus 2.0 arrays (Santa Clara, CA) according to manufacturer's instructions. Following hybridization, the probe arrays were washed and stained.
Scan protocol
The intensity of bound dye was measured with an argon laser confocal scanner (GeneArray Scanner, Agilent), and the stored images were analyzed using the GeneChip software Microarray Analysis Suite (MAS) 5.0 (Affymetrix, Santa Clara, CA).
Description
PGEF-Trained_1324 Gene expression data from a young healthy human subject.
Data processing
Raw intensity values of all samples were preprocessed and normalized by RMA using Bioconductor. Differentially expressed genes were tested by using an intensity-based Bayesian moderated t-statistic.
